黃連素抑制人類舌癌細胞株SCC-4生長的活體外和活體內的分子作用機轉之研究;Molecular mechanisms of growth inhibition on human tongue cancer SCC-4 cells by berberine：in vitro and in vivo

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(1)ύ୯ᙴᛰεᏢύ୯ᙴᏢࣴ‫܌ز‬റγፕЎ ࡰ Ꮴ ௲ ௤ Ǻ‫ߍ׵‬ী ௲௤ ӅӕࡰᏤ௲௤Ǻ݅ࡿ۪ ௲௤ ᗛඳӀ ௲௤. ፕЎᚒҞ ໳ೱન‫ڋ׭‬ΓᜪՁᕎಒझਲ਼ SCC-4 ғߏ ‫ࢲޑ‬ᡏѦ‫ࢲک‬ᡏϣ‫ޑ‬ϩηբҔᐒᙯϐࣴ‫ز‬ Molecular mechanisms of growth inhibition on human tongue cancer SCC-4 cells by berberineǺin vitro and in vivo. ࣴ‫ز‬ғǺՖ҉ᢕ. ύ๮҇୯ΐΜΖԃϖДΒΜΎВ!.

(2) ύ୯ᙴᛰεᏢύ୯ᙴᏢࣴ‫܌ز‬റγፕЎ. ࡰ Ꮴ ௲ ௤ Ǻ‫ߍ׵‬ী ௲௤ ӅӕࡰᏤ௲௤Ǻ݅ࡿ۪ ௲௤ ᗛඳӀ ௲௤. ፕЎᚒҞ ໳ೱન‫ڋ׭‬ΓᜪՁᕎಒझਲ਼ SCC-4 ғߏ ‫ࢲޑ‬ᡏѦ‫ࢲک‬ᡏϣ‫ޑ‬ϩηբҔᐒᙯϐࣴ‫ز‬ Molecular mechanisms of growth inhibition on human tongue cancer SCC-4 cells by berberineǺ in vitro and in vivo. ࣴ‫ز‬ғǺՖ҉ᢕ. ύ๮҇୯ΐΜΖԃϖДΒΜΎВ.

(3) Ҟᒵ ಃ΋കǵʳ ߻‫ ق‬...............................................................................................1 ಃΒകǵʳ Ў᝘௖૸ .......................................................................................5 ಃΟകǵʳ ‫׷‬਑ᆶБ‫ ݤ‬.................................................................................36 ಃѤകǵʳ ่݀ .............................................................................................67 ಃϖകǵʳ ૸ፕ .............................................................................................92 ಃϤകǵʳ ่ፕ .............................................................................................98 ୖԵЎ᝘ .....................................................................................................101 मЎᄔा .....................................................................................................113 ᖴᜏ .............................................................................................................116. i.

(4) კҞᒵ კ 3-1 ҁჴᡍ‫زࣴޑ‬೛ीࢎᄬკ............................................................36 კ 3-2 MMP ೬ᡏ‫ޑ‬ϩ‫݋‬კ......................................................................50 კ 3-3 ROS ೬ᡏϩ‫݋‬კ ............................................................................52 კ 3-4 ້ᚆη೬ᡏϩ‫݋‬კ.........................................................................54 კ 3-5 DAPI ࢉᏊᆶ DNA ‫ޑ‬ᗖ่............................................................55 კ 3-6 ᙯᅄ֨‫ޑ‬ϣ೽ಔԋ.........................................................................63 კ 4-1 ࢬԄಒझሺୀෳಒझӸࢲ౗ϐ่݀.............................................67 კ 4-2 ࢬԄಒझሺୀෳಒझຼයϐ่݀.................................................68 კ 4-3 ໳ೱનჹ SCC-4 ಒझਲ਼‫ ޑ‬MMP ϐ่݀ ....................................70 კ 4-4 ໳ೱનჹ SCC-4 ಒझਲ਼‫ ޑ‬ROS ϐ่݀ ......................................72 კ 4-5 ໳ೱનჹ SCC-4 ಒझਲ਼‫້ޑ‬ᚆηញ‫ܫ‬ϐ่݀............................73 კ 4-6 DAPI ࢉՅᢀჸ DNA ໾্ᆶಒझ঒Ϋ‫่݀ޑ‬...........................75 კ 4-7 Comet assay ᔠෳಒझ DNA ཞ໾‫่݀ޑ‬....................................76 კ 4-8 ໳ೱનჹ SCC-4 ಒझਲ਼‫ޑ‬ಒझຼයፓ௓ೈқ‫ޑ‬ᡂϯϐ߄౜..77 კ 4-9 ໳ೱનჹ SCC-4 ಒझਲ਼‫ ޑ‬mRNA ‫߄ޑ‬౜ᡂϯ .......................80 კ 4-10. ࢬԄಒझሺୀෳಒझӸࢲ౗ϐ่݀...........................................81. კ 4-11 ໳ೱનჹ SCC-4 ಒझਲ਼‫ڋ׭‬ᙯ౽‫่݀ޑ‬..................................82 კ 4-12. ໳ೱનჹ SCC-4 ಒझਲ਼‫ڋ׭‬ಒझߟΕ‫่݀ޑ‬..........................83. ii.

(5) კ 4-13 ໳ೱનჹ SCC-4 ಒझਲ਼࣬ᜢᙯ౽ೈқ‫ޑ‬ቹៜ............................85 კ 4-14 ໳ೱનჹ SCC-4 ဍዦ‫ޑ‬౦ᅿҜ౽෗ኳࠠ‫ޑ‬ჴᡍ೛ी................87 კ 4-15 ౦ᅿҜ౽෗୏‫ނ‬ኳࠠ‫ޑ‬ဍዦ߄౜ϐ่݀.....................................89 კ 4-16 ౦ᅿҜ౽෗୏‫ނ‬ኳࠠ‫ޑ‬ဍዦᡏᑈ‫ڋ׭‬౗.....................................91 კ 6-1 ໳ೱનჹ SCC-4 ಒझਲ਼ᇨวಒझ঒Ϋϐၡ৩............................98 კ 6-2 ໳ೱન‫ ڋ׭‬SCC-4 ಒझਲ਼‫ޑ‬ಒझᙯ౽‫ߟک‬Εၡ৩....................99 კ 6-3 ໳ೱન‫ڋ׭‬ᇘႵ SCC-4 ဍዦ‫ڋ׭ޑ‬౗......................................100. iii.

(6) ߄Ҟᒵ ߄ 3-1 ໳ೱનȐBerberineȑϐଛᇙ .........................................................43 ߄ 3-2 Ӹࢲ౗‫ ޑ‬PI ࢉᏊϐಔԋ ...............................................................47 ߄ 3-3 ಒझຼය‫ ޑ‬PI ࢉᏊϐಔԋ ...........................................................49 ߄ 3-4 Comet assay ϐ lysis buffer ଛᇙ .................................................57 ߄ 3-5 Comet assay ϐ Alkalin buffer ϐଛᇙ..........................................57 ߄ 3-6 Comet assay ϐ Tris buffer ϐଛᇙ ...............................................58 ߄ 3-7 ೈқ፦኱ྗࠔϐଛᇙ.....................................................................59 ߄ 3-8 SDS-PAGE ΠቫጤȐSeparation gelȑϐଛᇙϷಔԋ..................61 ߄ 3-9 SDS-PAGE ΢ቫጤȐStacking gelȑϐଛᇙϷಔԋ .....................61 ߄ 4-1 ճҔЇη߄ၲБԄຑ՗ DNA ‫ׇ‬ӈ ...............................................79. iv.

(7) ໳ೱન‫ڋ׭‬ΓᜪՁᕎಒझਲ਼ SCC-4 ғߏ ‫ࢲޑ‬ᡏѦ‫ࢲک‬ᡏϣ‫ޑ‬ϩηբҔᐒᙯϐࣴ‫ز‬. ࣴ‫ز‬ғ Ֆ҉ᢕ ࡰᏤ௲௤ ‫ߍ׵‬ী ௲௤ ύ୯ᙴᛰεᏢ ύ୯ᙴᏢࣴ‫܌ز‬. ! ! ҁፕЎϩࣁΟঁ೽ҽǴಃ΋೽ҽӃவ໳ೱનё೸ၸ caspase-8, -9 ‫ک‬ -3 ࿶җ AIF ᆶ EndoG ၡ৩ᇨวΓᜪՁᕎ SCC-4 ಒझਲ਼঒Ϋ‫ޑ‬ϩηբҔ ᐒᙯϐࣴ‫ز‬Ǵࣴ‫ز‬᛾ჴ໳ೱનፓ௓ SCC-4 ಒझਲ਼‫ޑ‬ಒझ঒Ϋࢂ೏ࢲ‫਼܄‬ ౢ‫ ( ނ‬ROS )ǵಈጕᡏǵCaspase-8, -9 ‫ک‬-3 ၡ৩‫܌‬ፓ௓ǶಃΒ೽ҽࢂ໳ೱ નё೸ၸ FAK, IKK, NF-țB, u-PA ‫ ک‬MMP-2 ‫ ک‬-9 ၡ৩‫ڋ׭‬ΓᜪՁᕎ SCC-4 ಒझਲ਼‫ޑ‬ᙯ౽ᆶߟΕբҔᐒᙯϐࣴ‫ز‬Ǵ่݀ᡉҢ໳ೱનӧΓᜪᡒ ‫ރ‬΢ҜՁᕎ SCC-4 ಒझਲ਼ૈӛΠፓ௓ uPA , MMP-2 ‫ ک‬-9 ‫߄ޑ‬ၲǴ೸ ၸ NF-țB ‫ ک‬MAPK ‫ޑ‬ፓ௓ၡ৩‫ک‬໳ೱન‫ޑ‬΋ঁཥ‫ޑ‬фૈǴё‫ڋ׭‬ൾ ‫܄‬ဍዦಒझ‫ߟޑ‬ΕૈΚǶಃΟ೽ҽ຾΋‫؁‬а໳ೱનჹΓᜪՁᕎ SCC-4 ಒ झਲ਼ӧᇘႵࢲᡏϣ‫ޑ‬ϩηբҔᐒᙯϐࣴ‫ز‬Ǵว౜໳ೱનӧ౦ᅿҜ౽෗୏ ‫ނ‬ኳࠠૈ‫ڋ׭‬ဍዦ‫ޑ‬ғߏǴ໳ೱનΨ೚ёբࣁՁᕎ‫ޑ‬Ⴃٛᛰ‫ނ‬Ǵ‫ޣ܈‬຾ ΋‫؁‬ԋࣁᖏ‫לޑ׉‬ᕎ‫ݯ‬ᕍᛰ‫ނ‬Ƕ. 1.

(8) ! ! ҁჴᡍ‫ޑ‬ϩηբҔᐒᙯ௶ॊӵΠǺ 1. ໳ೱનё೸ၸcaspase-8, -9‫ک‬-3 ࿶җAIFᆶEndoGၡ৩ᇨวΓᜪՁᕎ ಒझਲ਼SCC-4ಒझ঒Ϋ‫ޑ‬ϩηբҔᐒᙯϐࣴ‫ز‬ ! ! ෌‫ނ‬ϯᏢς࿶ዴᇡӧᕎੱ‫ݯ‬ᕍБय़ᏱԖወӧϯᏢႣٛ‫܈‬ϯᏢ‫ݯ‬ᕍ ‫ޑ‬фਏǴҁჴᡍଷ೛໳ೱનӧΓᜪՁᕎ SCC-4 ಒझਲ਼ஒԖ‫ל‬ᕎࢲ‫܄‬Ǵ ่݀ࡰр໳ೱન෧Ͽ SCC-4 ಒझਲ਼‫ޑ‬Ӹࢲ౗Ǵٗࢂ໒‫਼܄ࢲܭۈ‬ౢ‫ނ‬ ( ROS ) ‫ޑ‬ౢғ‫ک‬࿶җಒझϣ້ᚆη ( Ca2+ ) ‫ޑ‬ቚуǴ໳ೱનᇨวಒझ ঒Ϋࢂ‫ک‬ಈጕᡏಒझጢႝՏ ( MMP ) ‫ޑ‬෧ϿԖᜢǴΨ‫ک‬Bax/Bcl-2 К౗ ‫ׯ‬ᡂԖᜢǴಒझՅનC཮வಈጕᡏύញ‫ܫ‬ԶࢲϯΠෞ‫ޑ‬caspase-3ǶRealtime PCR ᡉҢ໳ೱન‫ڈ‬ᐟ caspase-8, 9 ‫ ک‬3 ಒझ঒ΫᇨวӢη ( AIF ) ‫ ک‬Endo G ‫୷ޑ‬Ӣ߄ၲǴҞ߻‫زࣴޑ‬᛾ჴ໳ೱનፓ௓ SCC-4 ಒझਲ਼‫ޑ‬ ಒझ঒Ϋࢂ೏ࢲ‫਼܄‬ౢ‫ ( ނ‬ROS )ǵಈጕᡏǵcaspase-3 ၡ৩‫܌‬ፓ௓Ƕࡌ ᝼໳ೱનΨ೚ёԵቾࣁ҂ٰࣴ‫ݯز‬ᕍՁᕎ٠ԋࣁԖ‫׆‬ఈ‫ݯޑ‬ᕍংᒧҔ ᛰǶ. 2. ໳ೱનё೸ၸFAK, IKK, NF-țB, u-PA ‫ ک‬MMP-2 ‫ ک‬-9ၡ৩‫ڋ׭‬ ΓᜪՁᕎSCC-4ಒझਲ਼‫ޑ‬ᙯ౽ᆶߟΕբҔᐒᙯϐࣴ‫ز‬ ! ! ֿᐟ䁙ࠠᠼᆢೈқྋ䁙চ໽ϯᏊ ( uPA ) ‫୷ک‬፦ߎឦೈқሇન ( MMPs ) ӧᕎੱᙯ౽‫ک‬Ոᆅཥғ‫ת‬ᄽࡐख़ा‫فޑ‬ՅǶ‫ ڋ׭‬uPA ‫ک‬ MMPs Ψૈ୼‫ڋ׭‬ᕎಒझ‫ޑ‬ᙯ౽‫ߟک‬ΕǶ໳ೱનࢂ໳ೱЬा‫่ޑ‬ᄬǴࢂ 2.

(9) ΋ᅿ౦ࠪ㭱ᜪғ‫ނ‬ᡵ‫ࠠޑ‬ԄǴൔ֋ࡰрӧόӕ‫ޑ‬ΓᜪᕎಒझԖ‫ל‬ᕎբ ҔǶฅԶ٠ؒԖൔ֋໳ೱનӧΓᜪՁᕎ SCC-4 ಒझਲ਼‫ޑ‬ᙯ౽‫ߟک‬Εբ ҔǶҁፕЎ‫ޑ‬ൔ֋่݀ᡉҢ໳ೱનӧΓᜪᡒ‫ރ‬΢ҜՁᕎ SCC-4 ಒझਲ਼ૈ ‫ڋ׭‬ᙯ౽‫ߟک‬ΕբҔǶ೭ঁբҔӧΓᜪᡒ‫ރ‬΢ҜՁᕎ SCC-4 ಒझਲ਼ࢂ೏ p-JNK, p-ERK, p-38, IțK ‫ ک‬NF-țB ߞ৲ၡ৩วғӧ‫ ڋ׭‬MMP-2 ‫– ک‬9 ‫܌‬ፓ௓Ƕ೭٤่݀ᡉҢ໳ೱનӧΓᜪᡒ‫ރ‬΢ҜՁᕎ SCC-4 ಒझਲ਼ ёӛΠፓ௓ uPA , MMP-2 ‫ ک‬-9 ‫߄ޑ‬ၲǴ೸ၸ NF-țB ‫ ک‬MAPK ‫ޑ‬ፓ ௓ၡ৩‫ک‬໳ೱન‫ޑ‬΋ঁཥ‫ޑ‬фૈஒё‫ڋ׭‬ൾ‫܄‬ဍዦಒझ‫ޑ‬ᙯ౽ϷߟΕ ૈΚǶ. 3. ໳ೱનჹΓᜪՁᕎ SCC-4 ಒझਲ਼ӧᇘႵࢲᡏϣ‫ޑ‬ϩηբҔᐒᙯϐࣴ ‫ز‬ ! ! ೚ӭ᛾ᏵᡉҢ໯१ύ‫ޑ‬Н݀‫ک‬ጫ๼ჹൾ‫܄‬ᕎ‫ک‬ठᕎ‫ނ‬፦‫ת‬ᄽٛៈ ‫ف‬ՅǶ໳ೱનࢂϺฅ෌‫ޑނ‬ಔԋǴ‫ٯ‬ӵλួǶ೚ӭࣴ‫ز‬ᡉҢ໳ೱનӧ೚ ӭᕎੱಒझਲ਼ᇨวಒझຼයଶᅉ‫ک‬ಒझ঒ΫǶҁፕЎЬा‫زࣴޑ‬ΨᡉҢ ໳ೱનӧΓᜪՁᕎ SCC-4 ಒझਲ਼ᇨวಒझ঒ΫǶԖᜢ໳ೱન‫ڋ׭‬Γᜪ Ձᕎ SCC-4 ಒझਲ਼‫ޑ‬ᇘႵࢲᡏϣ౦ᅿҜ౽෗୏‫ނ‬ኳࠠϝۘ҂Ԗൔ֋Ƕ SCC-4 ဍዦಒझ೏౽෗ΕᇘႵࢲᡏϣǴ‫ע‬ᇘႵϩಔ٠࿶ᛰ‫ނ‬ೀ౛Ǵ໳ೱ ન (10 డլ/ϦА/ᇘႵ*ǴDoxorubicin (4 డլ/ϦА/ᇘႵ)Ƕ೭٤ჴᡍᛰ ‫ࢂނ‬࿶ဎ๚ϣ‫؂‬ѤϺ‫ݙ‬৔΋ԛǴҔ 4 డլ/ϦА‫ ޑ‬Doxorubicin ‫ ܈‬10 3.

(10) డլ/ϦА‫ޑ‬໳ೱનೀ౛Ǵ่݀ᡉҢૈफ़եဍዦቹៜǴဍዦ‫ޑ‬ελǴӧ౦ ᅿҜ౽෗ᇘႵҔ 10 డլ/ϦА໳ೱનೀ౛ࢂܴᡉλ‫ܭ‬௓‫ڋ‬ಔǴ่݀ว౜ ໳ೱનӧ౦ᅿҜ౽෗୏‫ނ‬ኳࠠૈ‫ڋ׭‬ဍዦ‫ޑ‬ғߏǴ໳ೱનӧ҂ٰΨ೚ό ໻ஒёբࣁՁᕎ‫ޑ‬Ⴃٛᛰ‫ނ‬ǴࣗԿёૈԋࣁᖏ‫ݯ׉‬ᕍ‫ޑ‬Ԗਏᛰ‫ނ‬Ƕ. ᜢᗖӷ:ΓᜪՁᕎ SCC-4 ಒझਲ਼(Human tongue SCC-4 cancer cells)ǹ໳ ೱન (Berberine)ǹಒझ঒Ϋ (Apoptosis)ǹಒझຼය(Cell cycle)ǹCaspases ‫ޑ‬໺ሀ᜘(Caspase cascade)ǹಈጕᡏ(Mitochondria)ǹֿᐟ䁙ࠠᠼᆢೈқ ྋ䁙চ໽ϯᏊ (uPAǺUrokinase-type plasminogen activator)ǹᙯ౽ᆶߟΕ (Migration and invasion) ǹ ୷ ፦ ߎ ឦ ೈ қ 䁙 (MMPs Ǻ Matrix metalloproteinases )ǹਡᙯᒵӢη(Nuclear factor țBǺ NF-țB )ǹਡᙯᒵ ‫ڋ׭‬Ӣη(Inhibitory țBǺIțB). 4.

(11) ಃ΋ക ߻‫ق‬ ! ! Ѡ᡼Ӧ୔Ьा‫ޑ‬ԝΫচӢǴ٩Ᏽፁғ࿿ၗ਑಍ीǴ҇୯ΐΜϖԃϦ թ‫ޑ‬Ѡ᡼Ӧ୔ΓαЬाԝΫচӢύаൾ‫܄‬ဍዦଯۚಃ΋ՏǶѠ᡼Ӧ୔Ь ा‫ޑ‬ᕎੱԝΫচӢǴα๚ᕎ௨ӜΜεᕎੱԝӢಃϤՏǴԶ‫ځ‬ύ‫܄ت‬α๚ ᕎς៌ۚΜεᕎੱԝӢಃѤՏ. 1. ǴѠ᡼‫܄ت‬ᑡ஻α๚ᕎ‫ঁޑ‬ਢኧᡉҢр. Ԗ೴ԃቚу‫ޑ‬ᖿ༈Ƕӆу΢౜Ϟ‫཮ޗ‬ԃស่ᄬ೴ᅌ‫ז‬ೲԴϯǴᏃᆅᙴᕍ ሺᏔВᅌཥ⩧аϷᙴᕍ‫ೌמ‬В੻ගଯǴϝฅค‫ݤ‬ᗉխᕎੱ‫ુ࠶ޑ‬Ǵϸ‫ـ‬ ᑡᕎ౗όफ़ϸϲǶќѦ࿶ϯᏢᕍ‫ܫࢂ܈ݤ‬৔ᕍ‫ࡕݤ‬Ǵᕎಒझܰౢғ‫ל‬ᛰ ‫܄‬аϷऐ‫܄ڙ‬Ǵ‫׳‬ቚуΑᕎੱ‫ݯ‬ᕍ‫֚ޑ‬ᜤࡋǶӢԜǴࣴ‫ز‬ύᛰჹ‫ל‬ဍዦ ‫לޑ‬ᛰ‫܄‬Ϸᙯ౽Ǵ௦‫ࣽڗ‬Ꮲ‫ޑ‬ჴᡍኧᏵǴ‫ځڗ‬ୋբҔၨλ‫ޑ‬ᓬᗺǴճҔ ‫ځ‬ԖਏԋϩаϷբҔᐒᙯǴ໒วᕎੱႣٛᛰ‫ނ‬ǴஒԖշ‫ܭ‬ᕎੱ‫ޑ‬ႣٛϷ ‫ݯ‬ᕍǴ٠ёගϲੰΓ‫ޑ‬ғࢲࠔ፦Ƕ ! ! җ‫ܭ‬Ҟ߻୯ϣѦ٠ؒԖҺՖԖᜢ໳ೱન‫ڋ׭‬ΓᜪՁᕎ SCC-4 ಒझ ਲ਼‫ޑ‬Ў᝘ൔ֋ǴӢԜᒧ᏷໳ೱનࣁҁፕЎ‫زࣴޑ‬Ҟ‫ޑ‬Ǵ२Ӄа໳ೱન຾ ՉࢲᡏѦᕎಒझࣴ‫ز‬Ǵ௖૸໳ೱનբҔ‫ܭ‬ΓᜪՁᕎ SCC-4 ಒझਲ਼ࢂց ૈᇨวಒझ঒Ϋ‫ޑ‬ϩηբҔᐒᙯǶ຾΋‫؁‬௖૸໳ೱનբҔ‫ܭ‬ΓᜪՁᕎ SCC-4 ಒझਲ਼ࢂց‫ڀ‬Ԗғߏ‫ڋ׭‬Ϸᙯ౽‫ޑڋ׭‬բҔǴ٠௖૸‫ځ‬ᐒᙯǶ‫׳‬ ຾΋‫؁‬ӆ௖૸ᇘႵࢲᡏϣ‫ޑ‬Ձᕎಒझࢂց‫ډڙ‬໳ೱનғߏ‫ޑڋ׭‬ҔǴౢ ғ‫ڋ׭‬ဍዦελ‫ޑ‬ቹៜǴ٠௖૸‫ځ‬ϩηբҔᐒᙯǶ. 1.

(12) ! ! α๚‫ک‬αࠒᕎӧӄШࣚ‫؂‬ԃᑡ஻ΓኧຬၸΟΜ࿤ΓǴЪ‫܄ت‬ଯ‫ܭ‬ζ ‫܄‬30Ƕα๚ᕎ‫ޑ‬ᑡ஻‫ঁک‬Γ‫ੰࢉܰޑ‬੝፦࣬ᜢǴ‫ٯ‬ӵ੝ਸ‫୷ޑ‬Ӣ੝‫܄‬а Ϸό‫ޑؼ‬ғࢲБԄӢનǴ‫ٯ‬ӵ‫ྟܜ‬31. ǵ32. ǵᘓ཯33ǵഀଚ34Ǵ໯१ύલϿጫ. ๼‫ک‬Н݀35ǵલЮचᡀጱન‫ک‬ᆢғનA36ǵ༾ໆᔼᎦનό‫ى‬37 ᓐዦੰࢥགࢉ39όǵό‫ޑؼ‬α๚ፁғ‫࠻ک‬ϣ‫਻ޜ‬Ԧࢉ40. ǵ41. ǵ38. ǴΓᜪ٢. ฻Ӣન೿‫ת‬ᄽ. ख़ा‫فޑ‬ՅǴऩӕਔ‫ڀ‬ԖΒঁ‫׳܈‬ӭ‫ޑ‬೭٤Ӣનߡ཮ቚуᑡ஻Ձᕎ‫ޑ‬Ӓ ᓀ‫܄‬42. ǵ43. Ƕൔ֋ύࡰр‫کྟܜ‬സଚࢂᑡ஻α๚ᕎനЬाচӢ44. ǵ45. ǶਥᏵ. Չࡹଣፁғ࿿ൔ֋ࡰрӧѠ᡼‫ڟ‬ᝅᘓ཯ࢂᑡ஻α๚ᕎ‫ޑ‬΋ঁख़ा‫ޑ‬চ ӢǴ‫؂‬ԃεऊ‫؂‬΋ԭ࿤ΓαύԖ 9.6 ঁΓԝΫǶα๚ᕎ‫ݯࠠڂޑ‬ᕍхࡴ Ћೌǵ‫ܫ‬৔ጕ‫ݯ‬ᕍǵ‫ک‬ϯᏢᕍ‫ݤ‬ǹฅ‫ݯ‬ᘰ౗ࠅόࢂٗሶќΓᅈཀǴӢԜ ೚ӭࣴ‫ޣز‬ஒขᗺ‫ܫ‬ӧ‫ݯ‬ᕍα๚ᕎ‫ޑ‬ཥᛰว౜‫ک‬ཥᛰ‫ނ‬኱Ⴂ໒วࢂҞ ߻ख़ा‫ޑ‬ፐᚒǶ ! ! ࢬՉੰᏢ‫ک‬ჴᡍࣴ‫ز‬ᡉҢࢌ٤෌‫ނ‬ϯᏢӢηӧΓᜪᕎੱૈ୼‫ת‬ᄽ ӵӕϯᏢႣٛ‫܈‬ϯᏢ‫ݯ‬ᕍբҔ 46ǶԖ 121 ঁεӭኧ‫ޑ‬ೀБҔ٬Ҕ‫ܭ‬ᕎੱ ‫ޑ‬ೀ࿼Ǵ‫ࢂྍٰځ‬җ෌‫ڗނ‬ள 47Ƕ໳ೱનࢂϺฅ‫ޑ‬ғ‫ނ‬ᡵǴ᛾ჴࢂ‫ڈ‬ᔥ (Berberis vulgarisǵbarberry)ǵоߙယλួ(Berberis aquifoliumǵOregon grape)ǵ‫ڈؽ‬λᔥ(Berberis aristataǵtree turmeric)ǵቨโᛯ(Tinospora cordifolia)฻෌‫֡ނ‬ឦ‫ܭ‬໳ೱન‫ޑ‬ਥಳ༸Ъ‫ڀ‬Ԗࢲ‫܄‬ಔԋ 48Ƕ໳ೱન᛾ჴ ӧҜጥԖ‫ל‬๵ 49Ǵ‫਼ל‬ϯ 50Ǵ‫ל‬ว‫ ݹ‬51Ǵ‫ל‬ठᕎ฻ࢲ‫ ܄‬52Ƕ໳ೱન᛾ჴ. 2.

(13) ӧࢲᡏϣ‫ڀ‬Ԗ‫ל‬ว‫܄ࢲޑݹ‬. 53. ǶӧΓᜪ่ဉᕎಒझૈ‫ڋ׭‬. cyclooxygenase-2 ‫ޑ‬ᙯ᝿ࢲ‫ ܄‬54-56Ƕӧߚλယ‫ޤ܄‬ᕎ‫ڀ‬Ԗ‫ל‬ᙯ౽੝‫ ܄‬57Ƕ ໳ೱનΨ೏ว౜ӧว‫کݹ‬ठᕎ‫ޑ‬Γᜪಒझૈ‫ ڋ׭‬AP-1 ‫ޑ‬ख़ाᙯ᝿Ӣ ન‫ ڋ׭ک‬DNA ‫ܗ‬ᐆ౦ᄬ⅋ II ( DNA topoisomerase II ) 58ǶќѦԖൔ֋ ࡰр໳ೱનӧΓᜪқՈੰ U937 ಒझਲ਼‫ک‬ԴႵ໵Յનಒझዦ B16 ಒझ ਲ਼ᡉр‫ڀ‬Ԗಒझࢥનࢲ‫ ܄‬59ǶӧΓᜪ߄Ҝኬᕎ A431 ಒझਲ਼ 60 ‫߻ک‬ӈဏ ᕎಒझਲ਼. 61. ૈ‫ځڋ׭‬ғߏࢂᆶ caspases ࢲϯԖᜢǶӧ‫ޑॺך‬ჴᡍ࠻ς. ൔ֋ၸ໳ೱનӧ่ဉᕎಒझਲ਼ૈ‫ ڋ׭‬N-acetyltransferase ‫܄ࢲޑ‬. 62. Ƕ໳. ೱનӧΓᜪαࠒᕎ HSC-3 ಒझਲ਼ᇨวಒझຼයଶᅉᆶಒझ঒Ϋ‫ޑ‬ൔᏤ 63. Ƕ‫ڋ׭‬ႵՈᆅѳྖԼಒझ‫ࢲޑ‬ᡏѦቚ෗‫ک‬ᙯ౽. 63. Ƕՠᗋ҂Ԗᜢ‫ܭ‬໳ೱ. નӧΓᜪՁᕎಒझᇨวಒझ঒Ϋᆶ‫ڋ׭‬ΓᜪՁᕎ SCC-4 ಒझਲ਼‫ޑ‬ᙯ౽ ‫ߟک‬ΕϷӧՁᕎಒझ‫ࢲޑ‬ᡏ‫س‬಍ϣϯᏢᕍ‫ޑݤ‬բҔᐒᙯϐࣴ‫ޑز‬ൔ ֋Ƕ೭ঁࣴ‫ޑز‬Ҟ‫ࢂޑ‬ाΑှ໳ೱનӧΓᜪՁᕎ SCC-4 ಒझਲ਼ᇨวಒ झ঒Ϋ‫ޑ‬ϩηբҔᐒᙯᆶ໳ೱનӧΓᜪՁᕎ SCC-4 ಒझਲ਼ᇨวᙯ౽‫ک‬ ߟΕ‫ޑ‬ϩηբҔᐒᙯϷಃ΋ԛҾკࣴ‫ز‬ӧՁᕎಒझ‫ࢲޑ‬ᡏ‫س‬಍ϣϯᏢ ᕍ‫ޑݤ‬ϩηբҔᐒᙯϐࣴ‫ز‬Ƕ ! ! ҁፕЎࣴ‫่݀ز‬ᡉҢ໳ೱનё೸ၸ caspase-8, -9 ‫ک‬-3 ࿶җ AIF ᆶ EndoG ၡ৩ᇨวΓᜪՁᕎ SCC-4 ಒझਲ਼‫ޑ‬ಒझຼයଶᅉ٠ёᏤठಒझ ঒Ϋ 64 ᆶ໳ೱનё೸ၸ FAK, IKK, NF-țB, u-PA ‫ ک‬MMP-2 ‫ ک‬-9 ၡ৩. 3.

(14) ‫ڋ׭‬ΓᜪՁᕎ SCC-4 ಒझਲ਼‫ޑ‬ಒझᙯ౽‫ߟک‬Ε. 65. Ϸ໳ೱનჹΓᜪՁᕎ. SCC-4 ಒझਲ਼ӧᇘႵࢲᡏ‫س‬಍ϣ‫ڋ׭‬ဍዦ‫ޑ‬ғߏ 66Ƕ. 4.

(15) ಃΒക Ў᝘௖૸ 1.໺಍ύᙴჹ‫ܭ‬Ձᕎ‫ݤ࣮ޑ‬௖૸ ! ! ‫ך‬୯നԐԖᜢဍዦ‫૶ޑ‬ၩ‫ࣿࡾܭۈ‬Ꮿ୯ਔය‫ޑ‬໳ࡆϣ࿶ᡫኰǴ‫ځ‬ ύ൩Ԗҡ⡴ǵମੴǴϷဉዦǵโዦ‫ޑ‬ඔॊ2Ƕђжᠯǵ۟ǵᕎ೯ҔǶ ‫ځ‬ᖏ‫׉‬੝ᗺࢂֽ೽ဍ༧୲ฯǴଯեόѳǴҜՅόᡂǴ௢ϐό౽Ǵውឳ ࡕӵᙌ޸ҡᄱηǴՅ๋ൾૌǴੵภቃਗ਼Ǵόܰ‫ݯ‬ᘰǴࡺԖ๊ੱϐᇥǶ ӧᐕжЎ᝘ύዦᆶ۟ࢂϩ໒‫ޑ‬ǴᅬࣽЈள໣ύஒՁ๵ǵѨᄪǵ٢۟ǵ ๝۟ᙌ޸ᆀࣁѤε๊ੱǴ۟ࢂൾ‫ޑ܄‬Ǵዦӭኧࢂ‫ޑ܄ؼ‬3Ƕ ! ! ਥᏵҁੰ‫ޑ‬ᖏ‫׉‬੝ᗺǴᜪ՟‫ܭ‬ύᙴᏢϐȨУ۟ȩ ǵ ȨՁ๵ȩ ǵ ȨՁੳȩ Ǵ ឦ౜жᙴᏢ‫ޑ‬ՁᕎǶӧమǷଯ‫໋ޚ‬ϐȠᅬࣽЈள໣ȡ Ǵё‫ډـ‬ȨУ۟ȩ ‫ޑ‬Ԗᜢ૶ၩǴჹҁੰ‫ރੱޑ‬բΑ௶ॊǶӧమȠᙴ‫ߎے‬᠘ȡύǴ߾ჹ‫ځ‬ ੰӢǵੱ‫ރ‬ǵ‫ݯ‬ᕍ֡Ԗၨ၁ᅰ‫૶ޑ‬ၩǴ٠ܴዴࡰрȨՁ๵ȩ‫܄‬ൾǴఁ යಕϷᓍǵᓏԶр౜ฯ༧ϷႣࡕό‫ؼ‬฻੝ቻǶђжᙴᝤჹՁᕎ‫ޑ‬ඔॊ ೿ҔУ۟‫܈‬Ձ๵ᆶՁੳ‫ޑ‬ӜᆀǴϩॊӵΠǺ (1).ȠᅬࣽЈள໣ȡ4 ᇥǺ“⌭ဍँӵ‫ݱ‬ጪǴ‫ރ܈‬ӵᚊ߷ǴՁҁอᕭǴ όૈ՜๤Ǵ‫ق‬ᇟਔᅅૌతǶӆӢࡗ਻΢ፂǴ۹ฅ஝຋рՈόЗǴΦΦ ឳ‫ۯ‬У៱ǴջӜУ۟Ǵࣗ߾Ꭴဍ่ਡǴ୲ฯਔภǴҜՅӵதǴഗ೬΋ ᗺǴՅསόआǴઇࡕਔࢬૌНǴᆭӵ೬ᆟǴ‫ੱځ‬ᗨઇǴ୲ฯϝ߻όଏǴ Ԝ㵝ᆟውǴࣗԿ೸Ձ޵ဋǴ෯НᅅрǴࢂаΞӜᙌ޸۟ΨǶ”. 5.

(16) (2).Ƞᙴ‫ߎے‬᠘ȡ5ǴᆀՁੳǴჹ‫ځ‬ᖏ‫߄׉‬౜ǵੰำǵႣࡕ຾ՉΑಒጏ ‫ޑ‬ඔॊǴᒏǺ“Ձੳ‫ځ‬᛾നൾǴ߃ӵ‫ل‬Ǵԛӵ๵ǴᓐεဓλǴΞӜՁ ๵ǴੵภआឳคҜǴරᇸከख़ǶऩѨ‫ܭ‬ፓ‫ݯ‬Ǵаठ⌭ဍǴँӵ‫ݱ‬ጪǴ ‫܈‬Ԗ‫ރ‬ӵᚊ߷ǴՁҁอᕭǴόૈ՜๤Ǵ֫ᛖ໯१‫ق‬ᇟǴਔᅅૌతǶӆ Ӣࡗ਻΢ؑǴ۹ฅ஝຋ǴՈрόЗǴΦϐ‫ۯ‬Ϸ໨ᎤǴဍӵ่ਡǴ୲ฯ ԶภǴҜՅӵதǴ……ࣗ‫ޣ‬೸ՁऀဋǴ෯НᅅрǴ……ӢՁόૈᙯ୏Ǵ 㸦ଌ໯१Ǵࡺ‫؂‬१όૈк‫ى‬Ǵठगύ‫ޜ‬຀ǴԶੱ௃ቚख़ǴВᅌ૰௳Ǵ ԭό΋ғǶ”೭ᆶ౜жᙴᏢ‫ـ܌‬Ձᕎߚதௗ߈ǶԿ‫ځܭ‬วੰচӢǴӭ ೢϐЈ๠Оࢥ‫܌‬ठǶ” (3).ȠЀМോࣽઝਜȡ6 ᇥǺ“Ձ๵ឦЈ࿶ǴӭӢ਻ᢠԶғǶՁЗӵ๵ ‫ރ‬Ǵ‫܈‬ӵЕԸǴ‫ځ‬Յआ๋Ƕ” (4).Ƞკຏോࣽࡰඓȡ7 ᇥǺ“Ձੳϐ᛾ൾߚதǴЈ๠ОࢥᑈύѧǴ߃ ӵ‫ل‬εᅌӵ๵Ǵከख़රᇸ໯१֫Ǵࡗ߾஝ઇ೸ဋՁǴՍ‫ۯ‬໨ᓏਡ෷ܱǴ Ӝࣁⴔᘯ॥ᜤ‫ݯ‬ǴԭΓ஻Ԝԭ੃ΫǶ” (5).Ƞᅬᙴεӄȡ8Ǻ“ՁੳЈ๠ࢥОԋǴӵ‫ل‬ӵ๵ภឳआǴᅌऩ‫ݱ‬ጪ ᜤ໯१ǴᆟውΦᡂⴔᘯ॥Ƕ” ύᙴᇡ㵝Ձ㵝ЈϐभǴЈ໒ᘻ‫ܭ‬ՁǴՁҁឦЈǴЈે‫ܭس‬ՁਥǶ ѦགϤాǴΕ㚊ϯОǴϣ໾Ύ௃ǴᢠԶϯОǴठЈОᐥ౰Ǵൻ࿶΢‫؟‬ ‫ܭ‬ՁǶՁᜐឦ๠Ǵ๠ે๎‫ܭ‬Ձ਒Ǵኁࡘ໾๠Ǵ๠਻ᢠ่ёаϯО‫܈‬ၸ 6.

(17) १ٌ዗‫ݼ‬㿍ϐࠔǴύขᑈ዗ǴЈ๠ᢠОൻ࿶΢‫ܭ؟‬ՁǶ๝ϐࢭనр‫ܭ‬ ՁΠǴ๝຀ό‫ى‬Ǵ഍຀О‫ܮ‬Ǵ຀ОϣᐥǴՁѨ‫ځ‬ᎦǶคፕЈǵ๠ОࢥǴ ᗋࢂ຀Оϣ‫؟‬ǴࣣёठՁ΢தғውᅬǴуϐྟࢥ⡗੘Ǵ‫܈‬Уϐූਥූ ߷ߏය‫ڈ‬ᐟǴᏤठОࢥ࿀྿ϕ่‫ܭ‬ՁǴ࿶๎ߔ༞Ǵ਻Ո྿ᅉԶว㵝ҁ ੰǶё‫ـ‬ҁੰᆶЈ๠๝Ο࿶ஏϪ࣬ᜢǴЀ‫ࢂځ‬Ј๠ǴࡺȠᙴ‫ߎے‬᠙ȡ 5. ϓ“Ԝ᛾җЈǵ๠Оࢥ‫܌‬ठ”Ƕᖏ᛾аჴ᛾‫܈‬຀ύ਀ჴӭ‫ـ‬Ƕ. ! ! ӧύᙴᒣ᛾ፕ‫ݯ‬Бय़ǴёϩࣁϣܺᆶѦኦᛰΒᜪǴϩॊӵΠǺ (1) ϣܺᛰǺ ໳ೱᘞЈమОǴϯ྿ှࢥǶ຀‫ܺޣ‬ᘜ๠ ԐයЈО਻ᢠǴ߃ܺᏤ‫ه‬ණу໳ ෯ǴύයОࢥᐥ౰Ǵ዗ࣗ‫ܺޣ‬మథҒ៛໯ӝᘜ๠෯Ǵߡ▋‫ܺޣ‬ᘜ‫ؿ‬౦ ф෯ 5Ƕఁය਻Ո຀૰ࠠǴᒧҔံ਻ᎦՈǴှࢥණ่‫ޑ‬ᛰ‫ ނ‬9Ƕ (2) ѦҔᛰǺ a.ђᝤӭၩՁᕎ߃ଆҔ“ч৥Ϗᗺϐ”Ǵё੃ᕭԶཇǴమྛઝ໺ч৥ϏǺ พ㰤ࣳǵΓύқӚ 1.5 g Ǵґ΢ߙऱǵґ݊ǵ▋ᚊҠӚ 3 g Ƕ‫ݤڋ‬Ǻ Ҕ໼ሌᡞη‫ঁٿ‬Ǵᛰး‫ܭ‬ᡞϣǴஒα࠾ᝄǴѦҔᡶ‫ڰ࠾ݝ‬ǴаࣅОྪ आऊ 1 λਔǴ฻ࡑհ໒ᡞǴஒᛰ‫ڗ‬рǴΕ៭३ǵӇТӚ 1.3 gǴࣴಒ ҃ǴҔᅶ⿆‫ڈ‬ઇՁ๵ǴаϏϿ೚ᗺ΢Ƕ b.ϤઓΤ‫܈‬ఘ޸ᗺՁϏǴа፾ໆ࿼ੰᡂ೽Տ‫֖܈‬ϯࡕ໒Н‫܈‬໳ଚଌ ܺǴҔໆਥᏵதೕҔໆǶᅋαНБǺधୖ 30 gǴϖ७η 30 gǴξ‫ل‬ਥ 7.

(18) 30 gǴᓪဗ 30 gǴ૛‫ ًݞ‬30 gǴΕӇТϿ೚ྞ෯ǴжН֖ᅋǴ΋Вኧ ԛǶ ᎤΠԖဍ‫ޣނ‬ǺѦኦϯࢥණᆯ‫޳܈‬ᆺᆯ‫܈‬ᒸӦᛥᎉᑃᐚҊኦϐǴ‫܈‬Ƞᙴ ‫ߎے‬᠙ȡНዂᆯǺН०ԙࣳǵқ᭵ǵқ㐻ǵϖ७ηǵᢠߎǵ໢໳ǵ٢ ३ǴԯᎉፓᐚǴ៽ຠϐǶ c.ᐱ‫ف‬ጪѦኦ 10Ƕ. 2.౜жᙴᏢჹ‫ܭ‬Ձᕎ‫ݤ࣮ޑ‬௖૸ ! ! α๚ᕎࢂᓐᓍ೽ၨத‫ޑـ‬ൾ‫܄‬ဍዦϐ΋ǴԶՁᕎӧα๚ᕎύനࣁ ӭ‫ـ‬Ǵ౜жᙴᏢჹՁᕎ‫ੰޑ‬Ӣ᛾ჴᆶߏයၸࡋ֎ྟǵസଚǵᝅ१ᘓ཯Ǵ α๚ፁғৡǴ౦‫ߏނ‬ය‫ڈ‬ᐟǴՁℚጢख़ࡋқඬᆶआඬǴғ‫ނ‬ठᕎӢનǴ Ϸ‫ځ‬дӢનӵᒪ໺ǴᔼᎦжᖴምᛖǴ‫ܫ‬৔ጕ฻Ԗᜢ 11Ƕ ! ! ӧѠ᡼ᝅ१ᘓ཯Ꮴठα๚ᕎ‫ޑ‬วғ౗ε൯ቚуǴЀ‫ࢂځ‬ӧΒΜ ߻ǴѠ᡼࿶ᔮଆ‫ډ؁‬౜ӧǴΒΜԃ߻εໆᝅ१ᘓ཯‫ޣ‬ǴΒΜԃࡕԖࡐ ӭΓ൳Я‫ډ‬Αύԃ൩ள‫ډ‬α๚ᕎΑǴ೭ࢂࡐό۩‫ޑ‬Ǵၟ‫ځ‬д୯ৎК ၨǴ‫ځ‬д୯ৎ‫ޑ‬α๚ᕎ‫ޑ‬วғԃសࢂ 60-70 ྃǴԶѠ᡼‫ޑ‬ӳวғԃ សࢂ 40-45 ྃǶ ! ! Ձᕎ๊εӭኧ㵝ᡒ‫ރ‬ಒझᕎǴ΋૓ϩϯำࡋၨଯǴՁ߻ 2/3 Ձ ೽Տӭ㵝ᡒ‫ރ‬ಒझᕎǴ ࡕ 1/3 ӭ㵝ϩϯό‫ؼ‬ᕎǴϿኧࣁဏᕎǵరЃ ΢ҜᕎϷόϩϯᕎǵԺዦ‫ـش‬ǶՁᕎӭวғӧՁύ 1/3 ୁጔന㵝த 8.

(19) ‫ـ‬Ǵ‫ځ‬ԛ㵝ՁਥǵՁဎϷՁङǴՁӾന㵝Ͽ‫ـ‬Ƕᅐ‫ۯ‬ԿαۭϷᓏମǵ ՁᚓψǵࡧਲᡏǶՁᕎᙯ౽ཱུଯǴதวғӧԐයᓍరЃ่ᙯ౽Ǵᙯ౽ ౗㵝 60~80 % Ǵதӧ΋ୁǴӵੰᡂಕϷՁङ‫܈‬ຫၸՁᡏύጕǴёа ӛჹୁᓍరЃ่ᙯ౽Ƕ ! ! Ձᕎ೏ᘜឦ㵝α๚ᕎǴൾ‫܄‬ำࡋၨଯǴԐයࣁँрՁᡏ‫ޑ‬ဍ‫ނ‬Ǵ ‫׎‬ӵ‫ل‬ಈԶ፦ฯǴውឳࡕ‫׎‬ԋ୲ฯԶଯեόѳ‫ޑ‬ውᅬǶதวғԐයᓍ ೽రЃ่ᙯ౽Ǵఁයёᅐ‫ۯ‬αۭϷᓏମǴӵԖᝩวགࢉǴதวғቃภǶ ҁੰ੆ዎ‫܄‬மǴᕎዦёቶ‫ݱ‬ಕϷՁԼǴ٬Ձ‫ޑ‬ၮ୏‫ڙ‬ज़Ǵቹៜᇥ၉ǵ ຾१ϷտࠒǴ٠ԖӭໆࢬతǴૌᘸၳΓǶߟҍαۭ‫܈‬ຬຫύጕಕϷӄ ՁਔǴՁೀ‫ֹܭ‬ӄ‫ރۓڰ‬ᄊǴ㰗ғ஭αϷտࠒ֚ᜤ 12Ƕ ! ! ՁᕎੰำอǴว৖‫ז‬Ƕ߃යё‫ـ‬Ձ೽ғ΋λฯ่Ǵ‫׎‬ӵ‫ل‬ಈǴ೴ ᅌ‫׎‬ԋܴᡉဍ༧Ǵ٠ӧဍ༧ύЈ୔р౜ᜐጔ༾༾ໜଆ‫ޑ‬λውᅬǶ໒‫ۈ‬ ฯԶόภǴ‫܈‬ёߏεӵ๵ǴᓐεဓλǴёวғੵภǵᕧឳǴරᇸከख़Ǵ ੰᡂ೴ᅌӛు೽Ϸ‫ڬ‬ൎಔᙃᘉ৖Ǵӝ‫ٳ‬གࢉਔ㰗ғၨቃਗ਼‫ੵޑ‬ภǴё ӛӕୁ䏊य़೽‫ک‬Ը೽‫ܫ‬৔Ƕҁੰ੆ዎ‫܄‬மǴᕎዦёቶ‫ݱ‬ಕϷՁԼǴ٬ Ձ‫ޑ‬ၮ୏‫ڙ‬ज़Ǵቹៜᇥ၉ǵ຾१ϷտࠒǴ٠ԖӭໆࢬతǴૌᘸၳΓǶ ߟҍαۭ‫܈‬ຬຫύጕಕϷӄՁਔǴՁೀ‫ֹܭ‬ӄ‫ރۓڰ‬ᄊǴ㰗ғ஭αϷ տࠒ֚ᜤǶᕎዦውᅬёวғрՈǴ٠ёߟҍУ៱ǴࣗԿᓍᎤဍε୲ฯ ውᅬǶߟҍαۭϷᎤମǴё೸ՁऀဋǴ෯НࢬрǶఁයத٠วಔᙃᚯ 9.

(20) ԝǵрՈǵᔼᎦምᛖϷ֎Ε‫ݹޤ܄‬Ƕҁੰऊ 2/3 ‫ޑ‬஻‫ٳޣ‬วᓍరЃ่ ဍεǴᙯ౽రЃ่‫ޑ‬೽ՏаΒဎԼΠᓍు΢రЃ่നӭǴ‫ځ‬ԛࢂުछ ՁମԼа΢‫ޑ‬ᓍుύరЃ่‫ک‬ᎤΠǵ⯽ΠరЃ่Ǵᒿ๱ੰ௃຾৖Ǵᙯ ౽రЃ่ቚӭǴёጭ‫ډۯ‬ӕୁᙹମ΢‫܈‬ჹୁᓍ೽ǴఁයёวғՈՉᙯ ౽Ǵӭᙯ౽Կ‫ޤ‬೽ 13Ƕ ! ! Ձᕎࢂα๚ύൾ‫ࡋ܄‬നଯǴᙯ౽౗നε‫ޑ‬΋ᅿൾ‫܄‬ဍዦǴӢԜε ӭኧᕎੱ஑ࣽᙴৣ೿Ь஭ᆕӝ‫ݯ‬ᕍǶхࡴѦࣽЋೌϪନ‫ܫک‬৔‫ݯ‬ᕍᆶ ϯᏢ‫ݯ‬ᕍϷխࣝ‫ל‬ᕎᛰ‫ނ‬ᕍ‫ݤ‬ǶΒΜ΋Шइჹ‫ܭ‬ӄШࣚ‫ޑ‬ᕎੱੰΓᆶ ৎឦԶ‫ق‬Ǵᔈ၀ёаᆉࢂ΋ঁख़ࡌ‫׆‬ఈ‫ޑ‬ԃжǶӧ೭ࢤਔ໔္ǴԖ೚ ӭཥᛰԋф‫ࣴޑ‬ว΢ѱǴ೭൳ԃ྽ύǴѠ᡼‫ޑ‬ፁғ࿿Ψਡ঑Α೚ӭཥ ‫לޑ‬ᕎᛰǴж߄‫ޑ܄‬ᛰࠔхࡴၨԐයջ೯ၸऍ୯ᛰ‫ނ‬१ࠔᆅ౛ֽ (FDA) ਡ঑‫ޑ‬ϼѳࢩ๋ऽᎇ paclitaxelȐTaxolȑ67 Ϸऍ୯ᛰ‫ނ‬१ࠔᆅ ౛ֽ( FDA )ӧന߈೯ၸ‫ޑ‬ᐞࢪ๋ऽᎇ Docetaxel ᕎੱ‫ݯ‬ᕍཥᛰǴҔ‫ܭ‬ ‫ޤ‬ᕎǵ٢ᕎǵη৐ᓍᕎ֡ൔ֋Ԗ࣬྽ᕍਏ 68Ƕ. 3. ໳ೱ‫࣬ޑ‬ᜢЎ᝘௖૸ ! ! ໳ೱ㵝Лἇࣽ෌‫ނ‬໳ೱ Coptis chinensis FranchǵΟ‫ف‬ယ໳ೱ Coptis deltoidea CǷYǷCheng et Hsiao ‫܈‬໦ೱ Coptis teeta Wall ‫ޑ‬ଳᔿ ਥಳǶ. 10.

(21) (1).໳ೱǺғ‫ ܘੇܭ‬1000ɴ2000 ԯξӦஏ݅ύ‫܈‬ξ‫ك‬഍థೀǶഁғ‫܈‬ਭ ୻Ǵϩթ‫ܭ‬ଧՋǵ෫чǵ෫ࠄǵѤοǵ຦Ԁ฻ӦǶӧѤοܿ೽ǵ෫чՋ ೽Ԗၨεໆਭ୻Ƕ (2).Ο‫ف‬ယ໳ೱǺਭ୻‫ܭ‬Ѥοড়≒Ϸࢫ໡΋஥ੇ‫ ܘ‬1600ɴ2200 ԯϐ໔‫ޑ‬ ξӦ݅ΠǶ (3).໦ೱǺғ‫ ܘੇܭ‬1500ɴ2300 ԯϐ໔‫ޑ‬ଯξൣᔸ‫݅ޑ‬ጬΠǴഁғ‫܈‬ਭ ୻ǶϩթΪ໦ࠄՋч೽ϷՋᙒܿࠄ೽Ƕ Ȝ࡫ॣӜȝHuanglian ȜमЎӜ/܎΍ӜȝRHIZOMA COPTIDIS ȜձӜȝ‫ښ‬ೱǵ໡ೱǵ໦ೱǵοೱǶ Ȝٰྍȝ ᛰ‫ྍ୷׷‬ǺЛἇࣽ෌‫ނ‬໳ೱ(‫ښ‬ೱǵοೱ Coptis chinensis Franch) ǵΟ ‫ف‬ယ໳ೱ(Coptis deltoidea CǷYǷCheng et Hsiao)‫܈‬໦ೱ(Coptis teeta Wall) ‫ޑ‬ଳᔿਥಳǶϷ‫ځ‬дӕឦ߈ጔ෌‫ޑނ‬ଳᔿਥϷਥಳǴаѤο‫܌‬ ౢ‫ښޑ‬ೱ(οೱ)ࠔ፦ന٫Ƕ Ȝᛰ‫׷‬੝‫܄‬ȝ (1)Ƿ໳ೱǺਥಳӭᕠ‫ރ‬ϩ݄Ǵ៻Ԕϕ‫ܤ‬Ǵ‫׎‬՟ॹᚊП‫ރ‬Ǵಞᆀ“ᚊП ໳ೱ”Ǵൂ݄ᜪ༝ࢊ‫׎‬Ǵߏ 3ɴ6 ᙶԯǴ‫ޔ‬৩ 2ɴ8 డԯǶ߄य़Ԫ໳Յ ‫܈‬໳ලՅǴѦҜগပೀᡉआලՅǴಉᕫǴԖόೕ߾่࿯‫ރ‬ໜଆǴᠣਥ. 11.

(22) Ϸᠣਥූ୷ǴԖ‫ޑ‬࿯໔߄य़ѳྖӵಳาǴಞᆀ“ၸᐏ”Ǵ΢೽ӭූ੮ፃ ՅᡒယǴഗᆄத੮ԖූᎩ‫ޑ‬ಳ‫܈‬ယ࢓Ƕ፦୲ฯǴ‫ש‬ᘐय़ό᏾ሸǴҜ೽ ᐈआՅ‫܈‬སලՅǴ‫ࡋࠆځ‬ऊ㵝ъ৩‫ ޑ‬1ʏ3ǴЕ೽ᗲ໳Յ‫܈‬ᐈ໳ՅǴ ё‫ܫـ‬৔‫ރ‬દ౛Ǵᡎ೽आලՅǴԖਔύ‫ޜ‬Ƕ਻༾Ǵ‫ཱུښ‬धǶ (2)ǷΟ‫ف‬ယ໳ೱǺϩ݄ϿǴӭ㵝ൂ݄Ǵౣև༝ࢊ‫׎‬Ǵ༾៻ԔǴ4ɴ8 ᙶԯǶ‫ޔ‬৩ 5ɴ10 డԯǶၸᐏၨߏǶഗᆄԖϿ೚ූಳǶ፦ᇸԶฯǶ (3)Ƿ໦ೱǺౣևೱ੧‫ރ‬༝ࢊ‫׎‬Ǵӭ㵝ൂ݄Ǵ៻Ԕև㸃‫ރ‬ǴၨಒλǴ ߏ 2ɴ5 ᙶԯǴ‫ޔ‬৩ 2ɴ4 డԯǶ፦ᇸԶૄǶ Ȝ௦ԏ‫ک‬ᓯᙒȝࣿ‫ۑ‬௦ࡩǴନѐᠣਥϷ‫ݝ‬؅ǴଳᔿǴናѐූ੮ᠣਥǶ ȜϯᏢԋҽȝғ‫ނ‬ᡵᜪǺλᔥᡵǴ໳ೱᡵǴᛰਥᡵǴҘ୷໳ೱᡵǴЃ ଭߜ฻ǹ‫ځ‬дǺ᎑ǵᗔǵႇǵ؄ǵႉ฻คᐒϡનǶ Ȝ‫ښ܄‬ᘜ࿶ȝधǴൣǶᘜЈǵ๠ǵगǵ‫ط‬ǵᖌǵεဉ࿶ 15Ƕ Ȝҁ૛༼‫ق‬ȝ (1)Ƞઓၭҁ૛࿶ȡ 16Ǻ‫ښ‬धൣǶЬ‫ݯ‬዗਻ǴҞภǴ⃫໾Ǵ‫ݘ‬рǴ ܴҞǴဉⳏǴဎภǴΠฟǴ஁Γ഍ύဍภǶΦܺзΓό‫ב‬Ƕ (2)ȠӜᙴձᒵȡ 17Ǻ༾ൣǴคࢥǶЬ‫ݯ‬ϖᙒհ዗ǴΦΠ‫ⳏݧ‬ǵᖋ ՈǴЗ੃෰ǵεᡋǴନНǴճମǴፓगǴࠆဉǴ੻ᖌǴ‫ݯ‬αየǶ (3)Ƞҁ૛კ࿶ȡ 18Ǻ໳ೱ‫ݯ‬ҞБӭǴԶԲ‫ط‬ΤЀ‫ڻ‬౦Ƕᇂ౳Ҟϐ ੰǴࣣՈેᏉᅉ٬ฅǴࡺаՉՈᛰӝ໳ೱ‫ݯ‬ϐǶՈள዗߾ՉǴࡺ 12.

(23) ४዗ࢱΨǶ (4)Ƞҁ૛़ကȡ 19ǺϞΓӭҔ‫ݯ‬ฟǴᇂ୺аधᔿϐကǶΠ㚊ՠ‫ـ‬ ဉ຀ᅖ‫ݧ‬Ǵ༾՟ԖՈߡǴջҔϐǴ‫׳‬ό‫ޕ‬ЗǶΞό៝ൣ዗ӭϿǴ ՠаᅰᏊ㵝ࡋǴҗࢂӭठӒ֚Ƕऩ਻ჴ߃ੰǴ዗ӭՈฟǴܺϐߡ ЗǴϝόѸᅰᏊΨǶ‫܈‬຀ԶհǴ߾ό໪ܺǶ (5)Ƞ෯నҁ૛ȡ 20Ǻ਻ൣǴ‫ښ‬धǶ‫਻ࠆښ‬ᖓǴ഍ύ໚ΨǶϲΨǴ คࢥǶ ! ! ȠຝȡϓǺᘞЈОǴନ๠गύᔸ዗Ǵ‫ྠݯ‬ច⧋ЈǴᢠ዗ӧύ ขǴάάటӗǴЈΠฤᅈѸҔᛰΨǶҸඳ‫ݯ‬ΐᅿЈΠฤǴϖ฻ᘞ Ј෯ࣣҔϐǶѐᠣҔǶ ! ! ȠЈȡϓǺᘞЈ࿶ϐОǴ౳ኪ‫ه‬ဍϷፏየǴ໪ҔϐǶधൣ‫ޣ‬ Ь໚ԖᎩǴधаନϐǶӼຄǴ೯ൣ਱ǴᕍΠข຀Ǵ୲๝Ƕ ! ! ȠࣔȡϓǺଚ‫ݺ‬΢ՉǴଚ੆Չ΢ᓐǶ ! ! Ƞҁ૛ȡϓǺЬ዗਻ǴҞภ⃫໾‫ݘ‬рǴܴҞǶဉⳏဎภΠฟǴ ஁Γ഍ύဍภǶϖ᠌հ዗ǴΦΠ‫ⳏݧ‬ᖋՈǴЗ੃෰εᡋǴନНճ ମǴፓगࠆဉǴ੻ᖌǴᕍαየǶΦܺзΓό‫ב‬Ƕ ! ! ȠనȡϓǺΕЋϿ഍ǴधᔿǴࡺΕЈǴО൩ᔿΨǶฅᘞЈ‫ځ‬ ჴ๠ΨǴ㵝ηૈз҆ჴǴჴ߾ᘞ‫ځ‬ηǶ‫ݯ‬Ոٛ॥㵝΢٬Ǵ໳ೱ㵝 ύ٬ǴӦཾ㵝Π٬Ƕ. 13.

(24) ! ! ੇᙒ઒БǴзಖ‫ي‬όวඬየǺྞ໳ೱ΋αǴ‫ٽ‬ғ҂рᖂਔǴ ឲϐǴεᔈǶςрᖂឲϐǴඬᗨวǴҭᇸǶђБа໳ೱ㵝‫ݯ‬ฟϐ നǶ ! ! Ƞ़ကȡϓǺ‫ݯ‬ฟԖ༾ՈǴόё୺а໳ೱǶ㵝धᔿᏊǴ຀‫ޣ‬ ӭठӒ֚Ǵჴ‫ەޣ‬ҔϐǶ ! ! Ƞҁ૛ȡΞϓǺᓪମǵ౛ҡǵ໳᭷㵝ϐ٬Ǵൾ๸޸ǵ᭪޸ǵ ҎୖǵқᗲҜǴࣙීо޸Ǵയਜ਼ᓐǴှЃ‫ࢥل‬Ƕ (6)Ƞҁ૛ᆜҞȡ21Ǻ໳ೱεधεൣǴҔϐफ़ОᔿᔸǴύੰջ྽ЗǶ ଁёΦܺǴ٬๘ఠϐ਻தՉǴԶҵ‫ځ‬วғؑ‫ک‬ϐ਻Яǻ‫ݔ‬դ‫ق‬Ǻ ϖ‫ښ‬ΕगǴӚᘜ‫܌‬഻‫װ‬ǶΦԶቚ਻Ǵ‫ނ‬жϐதΨǶ਻ቚԶΦǴϽ ϐҗΨǶЦӇ‫ݙ‬ǺለΕ‫ط‬㵝ྕǴधΕЈ㵝዗ǴٌΕ‫ޤ‬㵝మǴយΕ ๝㵝ൣǴҒΕ๠㵝Կ഍ԶѤ਻ঋϐǴࣣቚ‫ښځ‬Զ੻‫਻ځ‬ǴࡺӚவ ҁ᠌ϐ਻㵝ҔǶ‫܌‬аΦܺ໳ೱǵधୖϸ዗ǴவОϯΨǴᎩ‫ࣣښ‬ฅǶ Φ߾᠌਻ୃയǴջԖୃ๊Ǵ߾ԖኪϽϐၰǶࢂа๊ಈܺራϐΓό ኪΫ‫ޣ‬Ǵคϖ‫ୃښ‬շΨǶ‫ݩ‬౳੯ҁ‫طܭ‬዗Ǵ‫ط‬ᆶЈ㵝η҆ǴЈОǴ ‫ط‬ҭОΨǴ๝‫᠌ې‬ΨǴΓ஻΋НόയΒОǴଁёΦܺधᛰǴ٬Ј Ԗ‫ୃ܌‬യǴࢂаО௱ОǴ‫ځ‬ёЯǻ ! ! ໳ೱǴ‫ݯ‬ҞϷฟ㵝ाᛰǶђБ‫ݯ‬ฟǴ३ೱΤȐЕ३ǵ໳ೱȑǶ ࠬೱණȐଳᖖҴȑ Ƕᡂ೯ΤȐ૤๹Ҵȑ Ƕᖖ໳ණȐғᖖҴȑ Ƕ‫੃ݯ‬෰. 14.

(25) Ҕଚᇃ໳ೱǶ‫ݯ‬ΠՈǴεᇁҴǶ‫ݯ‬αየǴಒٌҴǶࣣ΋հ΋዗Ǵ ΋഍΋໚ǴൣӢ዗ҔǴ዗ӢൣҔǴ։Խ࣬՘Ǵ഍໚࣬ᔮǴനளճ Бϐ֮Ǵ‫܌‬аԖԋфԶคୃയϐ্ΨǶ ླྀγᛁϓ:ѐЈᘻൾՈǴှܺᛰၸᏊྠ඀Ƕ (7)Ƞඳۢӄਜȡ 22Ǻ‫ښ‬εधǴ਻εൣǶ‫਻ࠆښ‬ᖓǴ؇ΨǴफ़ΨǴ फ़ύ༾ϲǴ഍ύ༾໚Ƕ஑‫ݯ‬ፏОǴОӧ΢Ǵ‫ݺ‬аଚǹОӧΠǴ‫ݺ‬ аูߡǶОԶჍ‫ݺޣ‬аᖖҊǹОԶҷ‫ݺޣ‬аᡶ෯Ƕӕֆ૤๹‫ݺ‬Ǵ ёаЗОภǹӕഋᏛβ‫ݺ‬ǴёЗ዗ᘞǶӕᱞჴҔǴё੃О๞ǹӕ Ϻ޸ણҔǴૈှྠ෰ǶӕЕ३ΤǴ‫ک‬ОᅉΠฟဎภǹӕֆ૤๹ΤǴ ‫ݯ‬ग዗ӗտለНǶᕴϐǴ‫܄ځ‬εൣǴࡺோѳ‫ط‬థՈǴ๘गమဉథ ᖌǴЗᡋ⻯ǴᘞЈନฤᅈǶ΢ё‫ݯ‬ӗՈἴՈǴΠё‫ݯ‬ဉߡआǶᕍ ஁Γ഍ЊဍภǴନλ‫ٽ‬१ᑈ዗ੳǴఠຄᙝǴ੃ൾየ㕢ဍǴନᔸ዗ ᢠ዗Ǵ๓‫ݯ‬О౳Ǵҭ੃౨ᅅǶှਜ਼ߕϐ዗ǴఠЃ‫ل‬ϐࢥǶฅ‫ځ‬๓ ᘞЈ๠ჴОǴ຀዗ӰҔǴѸठ਱໚Ǵࡺஊ‫ے‬በГǺ຀Զհ‫ޣ‬Ǵ཈ ϮᇸҔǶЦੇᙒГǺহДΦՈฟǴόҔ໳ೱǴ഍ӧϣΨǶඳۢГǺ Γϐ๠गǴ‫܌‬а౰ၩ࿤‫ނ‬Ǵวғ࿤‫ނ‬ǴҁຝӦԶឦβǶβཪ߾਻ ՉԶᔿǴβൣ߾਻ᏉԶᔸǴβᔿ߾ჴǴβᔸ߾ྖǴԜϺӦ໔όܰ ϐԿ౛Ƕ໳ೱϐधൣऩԜǴ‫܌‬аၸܺ᭷ೱ‫ޣ‬Ǵคό௳๠ǴԜ‫ځ‬ᔸ ྖǴҭԾܴᡉܰ‫ـ‬ǶᐱӢഏѶඳȠձᒵȡύԖፓगࠆဉϐ΋‫ق‬Ǵ. 15.

(26) Զቅ‫ݞ‬໔ൺ᛾ϐГǺፏधൣᛰӭ‫ݧ‬Ǵோ໳ೱǵ໳࢙‫܄‬հԶᔿǶӢ ठࡕШຎ㵝‫ـڻ‬Ǵคόᒏ໳ೱ‫܄‬ᔿԶࠆဉगǴΥ‫ݯ‬ᘞฟ‫ޣ‬Ǵ໒Ћ ߡࢂ໳ೱǴό‫ޕ‬໳ೱǵ໳࢙ϐᔿǴΪՖ‫ـ‬ϐǻ༖‫ڥ‬Ǽ΋‫ق‬ϐᙤǴ ࢬࢉऩԜǴᜤࢱऩԜǴ৺౛ൽΓǴವԜ㵝ࣗǶᗨГ໳ೱ‫ݯ‬ฟҭԖ ਏ‫ޣ‬ǴฅѸ‫ࢥځ‬࿪໚᠌Ǵ‫܈‬ӭᕵαဎǴᔸ዗㵝ฟ‫ޣ‬ǴΏ‫ە܌ځ‬Ƕ ЪΥаᕵစό࿯ԶՈ਻҅ம‫ޣ‬Ǵջ‫܈‬ᇤҔǴ҂ѸఠΓǴΦϐٕѐ ҭѸᅌཇǴԶᘜф໳ೱǴՖόёΨǻԜѦǴ߾Υаϡ਻ન১Ǵ໾ ๠஻ฟǴ‫܈‬ҁคОٕԶൣᔸ୏๠‫ޣ‬Ǵ‫ཱུੰځ‬ӭǴऩӰҔ໳ೱǴ߾ ๠๝В௳Ǵԭค΋ғǶΥ஻ฟԶԝ‫ޣ‬Ǵ౗җԜٰǴёόൣЈǻᎩ 㵝Ԝ‫ق‬ǴԶΓԖ҂Ѹߞ‫ޣ‬ǴӭаधᔿΒӷԖ҂ܴԸǴࡺէ‫ܭ‬Ƞ໺ ۸ᒵȡᒣ‫ݞ‬໔చύǴൺ၁‫ق‬ध‫ښ‬ϐ౛ǴаߩፁғϘ‫ޣ‬ӆ㵝ᜅ҅Ǵ பࢂߚளܴǴԶ҇ғԖ‫ױ‬ᒘ‫خ‬Ƕၰਜ‫ق‬Ǻܺ໳ೱҍ፜ԺǴзΓ‫ݧ‬ ᘞǶ (8)Ƞҁ૛ഢाȡ23ǺᘞОᔿᔸǶεधεൣǶΕЈᘞОǴЦੇᙒГǺ ᘞЈჴᘞ๠ΨǶჴ߾ᘞ‫ځ‬ηǶᙼ‫ط‬థՈǴΥ‫ݯ‬ՈǴٛ॥㵝΢೽ϐ ٬Ǵ໳ೱ㵝ύ೽ϐ٬ǴӦཾ㵝Π೽ϐ٬Ƕᔿᔸ໒ᢠǴှ෰ǴൂҔ ૈ‫੃ݯ‬෰ǶନྠǴ੻‫ط‬ᖌǴࠆဉगǴ੃Ј྿ǴૈѐЈᘻൾՈǶЗ หԠǶథЈǶ‫ݯ‬ဉⳏᘞฟǴߡՈГⳏǴԖ᠌ೱΤǶᔸ዗ᢠԶ㵝ฟǴ ໳ೱ‫ݯ‬ฟाᛰǶᏑα‫ޣ‬዗Ꮮ΢ขǴӕΓୖྞ෯‫ڡ‬ϐǴՠளΠࠒߡ. 16.

(27) ӳǶേ჏‫ق‬ГǺΠฟѸӃԠှ‫ځ‬ѦǴࡕፓ‫ځ‬ϣǶ२Ҕٌథаှ߄Ǵ ԛҔधൣа‫װ‬㚊Ƕ ȠᐒाȡϓǺࡕख़‫ە‬ΠǴဎภ‫کە‬Ǵ‫ي‬ख़‫ە‬ନᔸǴ ે۰‫ە‬ѐ॥Ǵ॥ٕϣ่‫ە‬ԠǴ‫ي‬հԾԠ‫ྕە‬ǴᖋՈ࿨ℚ‫ە‬ख़Ꮚа ᆃϐǶΠฟ‫ه‬ឦՈϩǴқឦ਻ϩǶᔎМГǺ߫ᒏ‫ه‬዗қൣ‫ߚޣ‬ΨǴ ೯բᔸ዗ೀ‫ݯ‬ǴՠԖཥΦ຀ჴϐϩǶฤᅈᔿᔸ໒ᢠǶҸඳ‫ݯ‬ΐᅿ ЈΠฤǴϖ฻ᘞЈ෯ࣣҔϐǶဎภǴమ዗ǶЈภҷఉǴЈᑈǶҞ ภ⃫໾ǴΓ٢੆ᗺ‫܈‬ӝᘜǵ‫ؿ‬฻ϩǴྞ෯዗ࢱǴණ዗ࢲՈǶ㕢ੴ የࣟǴፏภ᝚የǴࣣឦЈОǶܴҞȠ໺ߞБȡ ǺԲ‫ط‬΋‫ڀ‬Ǵ໳ೱ΋ ‫ٿ‬ǴཡΤǴΥࢂҞ੯ࣣ‫ݯ‬ǴӜԲ‫ط‬ΤǶ‫ۓ‬ᡋǴᙼ‫ط‬ǶЗԠှࢥǴ ନੳӕ፜‫ط‬ᇃ㵝ΤǶఠຄǶຄளध߾ҷǶ຀ൣ㵝ੰ‫ޣ‬࿣ҔǶΦܺ ໳ೱǵधୖϸ዗ǴவОϯΨǶܳࡪǺ‫ݹ‬΢բधǴ‫ښ‬धѸᔿǴᔿ߾ ዗‫خ‬ǶЪधൣ؇഍๘ఠǴҵ໾ғ‫ک‬ϐ਻ΨǶᗬҟГǺ໳ೱᆶԺਦ ӕՉǴૈҬЈ๝‫ܭ‬ഘ‫څ‬ǶਔࣔГǺ‫ݯ‬ฟҔ३ೱΤǹᖖೱΤҔ໳ೱǵ ଳᖖǹᖖ໳ණҔ໳ೱǵғᖖǹѰߎΤҔ໳ೱǵֆ૤๹Ƕ‫ݯ‬αየҔ ໳ೱǵಒٌǴЗΠՈҔ໳ೱǵεᇁǶ΋഍΋໚ǴൣӢ዗ҔǴ዗Ӣ ൣҔǶനள‫ڋ‬Бϐ֮Ƕ‫ݯ‬ЈОғҔǴ຀Оᎉ‫ݺ‬Ǵ‫ط‬ᖌО፜ᖌҊ‫ݺ‬Ǵ ΢ขОଚ‫ݺ‬ǴԖտለვᚇ฻᛾ǴҭԖӗለ‫ޣ‬Ӝ⑫ЈǴ‫ە‬໳ೱǵֆ ૤๹फ़О໒ᢠǶ⑫ǴॣᎉǶύขОᖖҊ‫ݺ‬ǹΠขНᡶН‫ݺߡู܈‬ǹ १ᑈО໳β‫ݺ‬ǹ‫ݯ‬ᔸ዗ӧ਻ϩǴֆ૤๹෯‫ݺ‬ǹӧՈϩଳᅊН‫ݺ‬ǹ. 17.

(28) ᗺ౳‫ه‬ǴΓ٢੆ǶਔࣔГǺፏ‫ݤ‬όᐱ㵝ϐЇᏤǴᇂٌ዗‫ځ‬ൣधǴ យൣ‫ځڋ‬ᔿ‫܄‬ǴҔ‫ޣ‬၁ϐǶ໳᭷ǵᓪମ㵝٬Ǵൾ๸޸ǵҎୖǵ⻄ ᡠǵқᗲҜǵࣙීоǵФጣǴ‫ג‬፜ԺǴਔࣔГǺБԖ᠌ೱΤǴ໳ ೱ፜‫ط‬ΤǴଁ‫ג‬ԺԶό‫᠌ג‬๜Яǻఠਜ਼ᓐǵЃ‫ࢥل‬Ƕ໳ೱᘞЈОǴ ՘аᓪᖌᘞ‫ط‬ᖌОǴқ‫ؿ‬ᘞ๠ОǴҡᆯᘞगОǴ‫҆ޕ‬ᘞ๝ОǴ໳ ࢙ᘞᆮાОǴЕ೯ᘞλဉОǴ໳᭷ᘞ‫ޤ‬ОǴఙη՘ϐǹᘞεဉОǴ ໳ೱ՘ϐǹਮचᘞ‫ط‬ᖌОǴ໳ೱ՘ϐǹᘞΟขОǴ໳᭷՘ϐǶᎄ ൚΋ГǺ዗ᢠ⧋ЈǴάάటӗǴҔ໳ೱኧϩࣗਏǶ (9)Ƞҁ࿶೹চȡ24ǺधൣǴคࢥǶ‫ݯ‬Ј᠌ОǴғҔǹ‫طݯ‬ᖌჴОǴ ፜ᖌҊ‫ݺ‬ǹ‫طݯ‬ᖌ຀ОǴᎉ‫ݺ‬ፃՅǹ‫ݯ‬΢ขОǴଚ‫ݺ‬ǹύขОᖖ Ҋ‫ݺ‬ǹΠขОǴᡶН‫ݺ‬ǹ਻ϩᢠ่‫ط‬ОǴྞֆ૤๹෯‫ݺ‬ǹՈϩ༧ ύҷОǴӕଳᅊ҃‫ݺ‬ǹ१ᑈОǴ໳β܏‫ݺ‬ǶှߕηǵЃ‫ل‬ǵᇸણ ࢥǴ‫ג‬፜ԺǶ ! ! ໳ೱ‫܄‬ൣ‫ښ‬धǴ਻ᖓ‫ࠆښ‬Ǵफ़ӭϲϿǴΕЋϿ഍ല഍ǶधΕ ЈǴൣയ዗Ǵ໳ೱǵε໳ϐधൣǴаᏤЈΠϐჴ዗ǴѐЈᘻൾՈǴ ҸඳΐᅿЈΠฤǴϖ฻ᘞЈ෯ࣣҔϐǶᘞЈ‫ޣ‬Ǵ‫ځ‬ჴᘞ๠Ǵჴ߾ ᘞ‫ځ‬ηΨǶΠฟगα຀዗αᏑ‫ޣ‬Ǵ໳ೱΓୖྞ෯Ǵਔਔ‫ڡ‬ϐǹӵ ӗӆ໯Ǵՠள΋‫ڡ‬ΠࠒߡӳǶፏधൣᛰӭᘞǴோ໳ೱǵ᭷ǵ࢙Ǵ ‫܄‬ൣԶᔿǴૈफ़ОѐᔸЗᘞฟǴࡺՈฟаϐ㵝։ǶϞΓՠ‫ـ‬ဉ຀. 18.

(29) ᅖ‫ݧ‬Ǵ༾՟ԖՈǴό៝ൣ዗ӭϿǴߡҔ໳ೱǴҗࢂӭठӒࢣǶԿ ‫ܭ‬຀հқฟǴϷӃᘞࡕฟϐ຀ൣ᛾ǴᇤҔठԝӭ‫خ‬ǶፏภᅬየǴ ࣣឦЈОǶ౳ኪ‫ه‬ဍǴภόё‫ה‬ǴҭឦЈОǴঋ਀‫ٕط‬Ǵॿ‫ە‬໳ ೱǵ྽ᘜǴ‫ݯ‬ฟϷҞ㵝ाᛰǶࡺȠҁ࿶ȡ२‫ݯق‬዗਻ҞภǴϷဉ ⳏဎภϐ஻Ǵ‫ڗ‬धᔿϐ‫܄‬ǴаమᓐҞǴ୲ဉगǴẮᔸ዗ΨǶ஁Γ ഍ύဍภǴҭࢂᔸ዗㵝஻ǴЀ‫ە‬аधᔿϐǶђБ‫ݯ‬ฟǴ३ೱΤҔ ໳ೱǵЕ३ǴᖖೱණҔଳᖖǵ໳ೱǴѰߎΤҔ໳ೱǵֆ૤๹Ƕ‫ݯ‬ ੃෰Ҕଚᇃ໳ೱǴ‫ݯ‬αየҔಒٌǵ໳ೱǴ‫ݯ‬ΠՈҔ໳ೱǵမᇁǶ ࣣࢂൣӢ዗ҔǴ዗ӢൣҔǴԶคୃയϐ্ǶฅधൣϐᏊǴύੰջ ЗǴଁё٬๘ఠϐзதՉǴԶҵғวؑ‫ک‬ϐ਻ЯǼᙴ࿶ԖΦܺ໳ ೱǵधୖϸ዗ϐᇥǴԜ‫܄‬ᗨൣǴ‫ښځ‬ԿधǴΕग߾Ӄᘜ‫ܭ‬ЈǴΦ ԶόςǴЈОୃയ߾዗ǴΏ‫ځ‬౛ΨǶ߈жঞࢬǴ഻Ҕ໳ೱ㵝మᏊǴ ਸό‫ޕ‬໳ೱᘞჴОǴऩ຀ОԶӰ‫׫‬Ǵϸ໾ύ਻Ǵ഍Оཇ଍΢ค‫ڋ‬ ‫خ‬Ƕࡺ഍຀ྠ዗Ǵ๠຀‫ݧ‬ᘞǴϖ‫׳‬๝‫ݧ‬Ǵ஁ΓౢࡕՈ຀ྠ዗Ǵλ ‫ٽ‬ร੶਻຀բᘞǴϷՉዀࡕ‫ݧ‬ᘞǴ٠ࣣ࿣ҔǶ (10)Ƞҁ૛‫੿؃‬ȡ 25ǺȘ‫ץ‬șεᘞЈОჴ዗Ƕ ! ! ໳ೱ஑ΕЈǴঋΕဉǵगǵ๠ǶεधεൣǴᏵਜ‫܌‬ၩ‫ݯ‬фǴ ഢཱུ߄๱ǴЪаȠձᒵȡύԖࠆဉग΋ᇟǴϕ㵝໺ኞǴаԿ‫ܭ‬ϞǴ ᙤЀವၬǴຢ্คጁǶ␢‫ޕ‬໳ೱЗឦᘞЈϐࠔǴନᔸϐ‫ښ‬Ƕӳђ. 19.

(30) ГǺ໳ೱधᔿǴधΕЈǴЈ൩ᔿǴᘞЈ‫ޣ‬Ǵ‫ځ‬ჴᘞ๠ΨǶჴ߾ᘞ ‫ځ‬ηΨǶջϓဉⳏૈЗǴαଳૈନǴฤᅈဎภૈ੃Ǵ㕢ੴየᅬૈ ཇǴ‫ط‬຀ૈᙼǴᆶϻ஁Γ഍ᇑǴλ‫ٽ‬ੳᑈǴ٠О౳‫ه‬ภǴӗՈǵ ἴՈǵፏࢥ฻ੱǴคόҗԜፓ‫ݯ‬ǴҭՖವόӢᔸ዗ОଏԶ‫ق‬Ǵଁ ‫ܭ‬ᔸନОଏϐѦǴۘԖ‫ݯ‬ਏϐ๱ࠌǶϡનГǺ໳ೱ‫ځ‬ҔԖϤǴᘞ Ј᠌ОǴ΋Ψǹѐύขᔸ዗ǴΒΨǹፏየѸҔǴΟΨǹѐ॥ᔸǴ ѤΨǹ‫ه‬౳ኪวǴϖΨǹЗύ೽‫ـ‬ՈǴϤΨǶԙ᎜ՎГǺΠฟग α዗Ꮡα‫ޣ‬ǴҔ໳ೱΓୖྞ෯ǴಖВ‫ڡ‬ϐǴӵӗǴӆம໯Ǵՠள ΋‫ڡ‬ΠࠒߡӳǶቅֹનГǺђБа໳ೱ㵝‫ݯ‬ฟϐനǴᇂ‫ݯ‬ฟோ‫ە‬ ٌधൣᛰǴٌૈวණǴ໒೯ᢠ่ǴधૈᔿᔸǴൣૈയ዗Ǵ٬਻࠹ ѳԶςǶፏधൣᛰӭ‫ݧ‬Ǵோ໳ೱǵ໳࢙‫܄‬հԶᔿǴૈफ़ОѐᔸԶ ЗᘞǶГǺϞΓӭҔ໳ೱ‫ݯ‬ฟǴᇂ୺аधᔿϐကǴΠ㚊ՠ‫ـ‬ဉ຀ ᅖ‫ݧ‬Ǵ༾՟ԖՈǴߡջҔϐǴΞό៝ൣ዗ӭϿǴோటᅰᏊǴҗࢂ ӭठӒ֚ǶਔࣔГǺ໳ೱεधεൣϐᛰǴҔϐफ़ОᔿᔸǴύੰջ ྽ЗǴଁёΦܺǴ٬๘ఠϐзதՉԶҵ‫ځ‬ғวؑ‫ک‬ϐ਻ЯǶછᢀ ᆶൈ‫׆‬ဃፕ໳ೱਜϓǺᆪϦа౳੯ራ໳ೱԿΜኧ‫ٿ‬ซόςǴࢣό ёΨǶᙴ࿶ԖΦܺ໳ೱǵधୖϸ዗ϐᇥǴԜᗨεൣǴ‫ښځ‬ԿधǴ Εग߾Ӄᘜ‫ܭ‬ЈǴΕԶόςǴЈОୃയ߾዗ǴΏ‫ځ‬౛ΨǶ᳘Ԗ๠ ໚ન১ǴӢԜ΋‫ࢬق‬ኞǴԶёࡢ㵝தܺ‫ޣ‬ЯǶϞΓ΋‫ـ‬ОᐥǴό. 20.

(31) ፕࢂൣࢂ዗Ǵࢂ຀ࢂჴǴᇷа‫׫ڗ‬Ǵаठୃയຢ஻Ǵས‫ڙ‬Ͻ‫ש‬Ǵ ਸ൐ኼெǶਔࣔГǺ໳ೱ‫ݯ‬ҞϷฟ㵝ाᛰǶђБ‫ݯ‬ฟǴ३ೱΤҔ ໳ೱǵЕ३ǹᖖೱණҔଳᖖǵ໳ೱǹᡂ೯ණҔ໳ೱǵ૤๹ǹᖖ໳ ණҔ໳ೱǵғᖖǶ‫੃ݯ‬෰Ҕଚᇃ໳ೱǹ‫ݯ‬ҷපҔଚฆ໳ೱǹ‫ݯ‬Π ՈҔ໳ೱǵεᇁǹ‫طݯ‬ОҔ໳ೱǵ૤๹ǹ‫ݯ‬αየҔ໳ೱǵಒٌǴ ࣣࢂ΋հ΋዗Ǵ΋഍΋໚Ǵൣ዗ϕҔϐཀǴԶคୃയϐ্Ƕ؋ܳ ГǺ໳ೱᘞЈОǴ՘аᓪᖌᘞ‫ط‬ᖌОǴқ‫ؿ‬ᘞ๠ОǴҡᆯᘞगОǴ ‫҆ޕ‬ᘞ๝ОǴ໳࢙ᘞᆮાОǴЕ೯ᘞλဉОǶ໳᭷ᘞ‫ޤ‬ОǴఙη ՘ϐǹᘞεဉОǴ໳ೱ՘ϐǶਮचᘞ‫ط‬ᖌОǴ໳ೱ՘ϐǹᘞΟข ОǴ໳᭷՘ϐǶᙇࡪǺਮचᘞОǴЗ൩‫ط‬ᖌٕᢠԶ‫ق‬ǴऩϣჴО ҔԜǴཇቚ‫خ্ځ‬Ǵόёό‫ޕ‬Ƕ (11)Ƞҁ૛࿶ှȡ26Ǻ໳ೱ਻ൣǴ‫ޚ‬ϺоൣϐН਻ǴΕ‫ى‬Ͽ഍๝࿶ǹ ‫ښ‬धคࢥǴளӦࠄБϐО‫ښ‬ǴΕЋϿ഍Ј࿶Ƕ਻‫ॿښ‬फ़Ǵ഍ΨǶ ! ! ‫ځ‬Ь዗਻ҞภΨǴЈЬОǴО਻዗ǴЈੰް‫ط‬Ǵ‫ط‬໒ᘻ‫ܭ‬Ҟ ΨǴ໳ೱधൣǴ‫܌‬аమОΨǶЋϿ഍ϐ҅ેǴр‫ܭ‬य़ǴӝҞϣ⃫Ǵ ЋϿ഍㵝ЈОǴО౰Ǵ߾Ј‫࡚س‬Զఽрǹ⃫໾‫ޣ‬ǴࣣЈОǴ໳ೱ మЈǴ‫܌‬аЬϐǶჴ߾ᘞ‫ځ‬ηǴЈ‫ޣ‬Ǵ‫ط‬ЕϐηΨǴమЈ߾‫ٕط‬ ᘞǴ‫܌‬аܴҞΨǶ ! ! εဉ㵝۪ߎϐ๜ǴЈО४ϐǴ߾ࢭనϯԋᖋՈǴภԶΠฟ‫خ‬Ǵ. 21.

(32) ‫ځ‬Ьϐ‫ޣ‬ǴൣаమОǴधаᘞ዗ΨǶ ! ! чБ໵ՅǴΕ೯‫ܭ‬๝Ǵ໒ᘻΪΒ഍Ǵ஁Γ഍ύǴΏ๝ᘻΨǴ ዗౰߾ဍǴဍภ‫ޣ‬О౰ΨǴ໳ೱΕ๝ǴൣधమОǴ‫܌‬аЬϐǶ ! ! ‫ځ‬ΦܺзΓό‫ޣב‬ǴΕЈమОǴОమ߾ЈܴǴૈ૶ᏫΨǶ (12)Ƞઓၭҁ૛࿶᠐ȡ27Ǻ໳ೱ਻ൣǴ‫ޚ‬ϺоൣϐН਻ǴΕ‫ى‬Ͽ഍ ๝࿶ǹ‫ښ‬धคࢥǴளӦࠄБϐО‫ښ‬ǴΕЋϿ഍Ј࿶Ǵ਻НԶ‫ښ‬НǴ ΋‫ނ‬ӕॿǴࡺૈନНО࣬໶ǴԶ㵝ᔸ዗ϐੰǶ ! ! ‫ځ‬ϓϐ዗਻‫ޣ‬Ǵନ΋Ϫ਻ϩϐ዗ΨǶҞภ⃫໾ఽрόܴǴࣣ ᔸ዗ӧ΢ϐੰǹဉⳏဎภΠฟǴࣣᔸ዗ӧύϐੰǹ஁Γ഍ύဍภǴ 㵝ᔸ዗ӧΠϐੰǴ໳ೱନᔸ዗Ǵ‫܌‬аЬϐǶ ! ! ΦܺзΓό‫ޣב‬ǴधΕЈǴջૈံЈΨǶฅध㵝Оϐҁ‫ښ‬Ǵ а‫ښځ‬ϐधԶံϐǹԶൣૈയОǴջа‫ځ‬ϐൣԶᘞϐǶίђ୤Ҹ ඳளȠҁ࿶ȡϐઝǴ Ƞߎ჆ȡ‫ݯ‬Ј਻ό‫ى‬ԶӗՈ‫ޣ‬Ǵ‫ڗ‬ϐаံЈǹ ໾ൣൣ዗ϕ่ЈΠǴԶฤᅈ‫ޣ‬Ǵ‫ڗ‬ϐаᘞЈǹല഍ϐ዗Ǵ਻ናЈ ‫ޣ‬Ǵӝаਜ਼ఘǹΠฟࡕख़‫ޣ‬Ǵӝақᓐશ฻‫ݤ‬Ǵ੿ߞԶӳђϐε ဃΓΨǶ (13)Ƞҁ૛ϩ࿶ȡ28ǺεधεൣǶΕЈᘞОǴᙼ‫ط‬థՈǴᔿᔸ໒ᢠǴ ૈ੃ЈᘻൾՈǴҭᘞ๠ОǶଚ‫ݯݺ‬΢ขОǴᖖҊ‫ݯݺ‬ύขОǴН ‫ݯݺ‬ΠขО. 16. Ƕ. 22.

(33) ᕴԶ‫ق‬ϐǴ໳ೱ‫ښ‬ध‫܄‬ൣǴᘜ࿶Ј Ε‫ ط‬Εᖌ Ε๠ Εग Εεဉ฻࿶Ǵ ឦమ዗ᔿᔸᛰǴҁჴᡍ჋၂ճҔ໳ೱ‫ޑ‬ᛰ‫ނ‬੝‫܄‬٬Ҕ‫ܭ‬Ձᕎಒझ ‫زࣴޑ‬Ƕ. 4. ໳ೱનȐBerberineȑ‫ޑ‬Ў᝘ӣ៝ ȜᛰࠔӜȝ໳ೱનȐBerberineȑ ȜձӜȝλួᡵ ȜϯᏢӜȝ5,6-dihydro-9,10-dimethoxybenzo[g]-1,3-benzodioxolo[5,6a] quinolizinium [CAS] Ȝ.ϩηԄȝ[C20H18NO4]+ Ȝϩηໆȝ336.37 ȜϯᏢ่ᄬԄȝ໳ೱનȐberberineȑ‫ޑ‬ϯᏢ่ᄬ‫ރ׎‬. Ȝ‫ނ‬౛ϯᏢ‫܄‬፦ȝλួᡵࣁ໳Յણ҃Ǵᅙᗺ 145ʚǴෞᚆ‫ޑ‬λួᡵૈ ጗ᄌྋှ‫ܭ‬НύǴჹհΌᎇྋှࡋόεǴՠܰྋှ‫ܭ‬዗НϷ዗ΌᎇǴ ᜤྋ‫ܭ‬शǵෛү‫ک‬Ч✉ 69Ƕ 23.

(34) ȜЎ᝘ϟಏȝ ! ! ໳ೱનΞᆀλួᡵǴ㵝Лἇࣽ໳ೱឦ෌‫ނ‬໳ೱǵΟ‫ف‬ယ໳ೱϷ໦ ࠄ໳ೱ‫ޑ‬ਥಳύග‫ޑڗ‬౦ࠪ㭱ᜪғ‫ނ‬ᡵǴதҔ‫ځ‬ᡶለᡶǶӧᛰ౛բҔ ΢‫ڀ‬Ԗ‫ל‬༾ғ‫ނ‬Ϸ‫ל‬চᙝ‫ޑ‬բҔǶ‫ל‬ᕎբҔǶჹઓ࿶‫س‬಍‫ޑ‬բҔǴӧ λႵٛᑇၮ୏‫܄‬చҹϸ৔ჴᡍύǴ໳ೱનλᏊໆૈߦ຾໚‫܄‬చҹϸ৔ ‫׎ޑ‬ԋǴԶεᏊໆ߾‫ۯ‬጗‫׎ځ‬ԋǴ٠ё٬చҹϸ৔ϩϯόӄǶჹѦ‫ڬ‬ ઓ࿶ሀ፦‫ޑ‬ቹៜǴӧ᏾ᡏၮ୏‫ک‬ᚆᡏᏔ‫۔‬΢Ǵ໳ೱનჹΌㄽᖌᡵ‫ڀ‬Ԗ Ꮚໆ٩ᒘ‫܄‬ᚈख़բҔǴ΋૓೿ࢂλᏊໆቚуΌㄽᖌᡵ‫ޑ‬բҔǴԶεᏊ ໆ෧১ϐǶჹЈ᠌‫ޑ‬ቹៜǴ໳ೱનեᐚࡋૈᑫᏟᒗᚆᡏЈ᠌Ǵ٠ቚу ߷ેՈࢬ 20%ɴ40%Ǵ1×105 ߾߄౜ࣁ‫ڋ׭‬Ǵՠ 1×10-4 ΨόठଶЗཛ ୏ǶჹՈᆅϷՈᓸ‫ޑ‬ቹៜǴ໳ೱનᓉે‫ݙ‬৔‫ܭ‬ഞᎈХǵᒗǵεႵϷό ഞᎈεႵԖܴዴ‫ޑ‬फ़ᓸբҔǶჹѳྖԼᏔ‫ޑ۔‬ቹៜǶჹಔᙃжᖴ‫ޑ‬ቹ ៜǴ໳ೱનૈफ़եλႵ‫ط‬ǵတϬዀ‫઻਼ޑ‬ǴЪբҔமࡋᆶᏊໆ࣬ѳՉǶ ჹՈన‫ޑ‬ቹៜǴමว౜໳ೱનૈ٬आՈౚᕭλԋᗭಈ‫ރ‬Ǵ٠‫ڋ׭‬қՈ ౚᡂ‫׎‬ၮ୏Ǵ෧Ͽύ‫܄‬Ϸ༐ለ‫܄‬ಒझԶቚуరЃಒझϷൂਡಒझ 15Ƕ ᖏ‫׉‬΢‫ޑ‬фҔឦ‫ܭ‬మ዗ǵှࢥϐբҔǵё‫ݯ‬ᕍಒ๵‫܄‬ฟ੯ǵ໾ൣǵࢬ Չ‫܄‬တૉᡎጢ‫ݹ‬ǵεယ‫ݹޤ܄‬ǵฎआ዗ǵқോǵ‫ޤ‬ᖋဍǵᖋૅǵ‫ޤ‬қ Յ‫ۺ‬੧๵ੰǵውᅬ‫่܄‬ဉ‫ݹ‬ǵ่ဉλ೓ᠼЛᙝੰǵଯՈᓸǵϯᖋ‫܄‬ύ Ը‫ݹ‬ǵѲМఎ๵ੰǵԭВࠏǵᐨ໾ǵϯᖋ‫܄‬གࢉǵ‫ع‬຋ǵମᜢ࿯่ਡ. 24.

(35) ᝠၰǵ๺ᕭ‫܄‬ሷ‫ݹ‬ǵࠒ৙‫ݹ‬ǵ࡚‫ࡧ܄‬ਲᡏ‫ݹ‬ǵ΢ᓏᝠ‫ݹ‬ǵਥᆅ੯஻ǵ α๚ᓏय़೽‫ੱݹ‬ǵҜጥགࢉ‫ੱݹ܄‬ǵᔸ੶ǵӭࠠ‫܄‬ᅖр‫܄‬आඬǵОᐭ ໾ǵٛ‫ݯ‬࿂੶฻ᖏ‫׉‬բҔ 29ǶӧғϯϷᇙᛰᏢሦୱύԖ࣬྽‫ޑ‬ᔈҔǶ ໳ೱનҭ೏ว౜Ԗ‫ל‬ᕎբҔǴӧη৐ᓍᕎ HeLa ಒझਲ਼ϷқՈੰ L1210 ಒझਲ਼ύᇨว DNA ‫ܗ‬ᐆ౦ᄬ䁙ȐENA topoisomeraseȑࢥ্೷ ԋಒझ‫ޑ‬঒Ϋ 70ǶѬҭૈᙖҗ‫ ڋ׭‬AP-1 ‫܄ࢲޑ‬Զ‫ڋ׭‬Ϸፓ௓రЃ่ ᕎಒझਲ਼‫ޑ‬ᙯ౽ 71Ƕ໳ೱન߄౜ࢲᡏϣϷࢲᡏѦ‫לޑ‬ว‫ݹ‬բҔ. 72. Ƕ໳. ೱનϷ‫ځ‬ᜪ՟‫ނ‬Ԗ‫ל‬ဍዦբҔǴЬाࢂ೯ၸ‫ڋ׭‬ዦಒझ‫֎ڥ‬ǵಒझ❥ ᩫ‫ک‬ਡለ‫ޑ‬ғ‫ނ‬ӝԋᇨᏤ঒ΫբҔԶวචբҔ 73Ƕ໳ೱનೀ౛ࡕǴ‫ڀ‬ (DNA ӝԋ߻ය)S ය‫ޑ‬ဍዦಒझК‫ٯ‬εε෧ϿǴಒझߔᅉ‫ ܭ‬G0 ‫ک‬ G1 යǴ٬ಒझቚ෗‫ڬ‬ය‫ߏۯ‬வԶᏤठಒझቚ෗෧ᄌ 74Ƕ໳ೱનҭё ᇨᏤဍዦಒझ঒ΫǴҔ໳ೱનೀ౛Γगᕎ MGC-803 ಒझਲ਼ࡕǴࢉՅ ‫ک‬চՏ҃ᆄ኱૶‫ݤ‬຾Չᔠෳว౜ MGC-803 ಒझਲ਼և‫ޑࠠڂ‬঒Ϋ੝ ቻ 75Ƕ໳ೱનᆶ‫ל‬ᕎᛰ CPT-11 ᖄӝҔ‫ܭ‬λႵ‫ޤ‬ᕎ౽෌ዦǴ‫ځ‬ဍዦғ ߏ‫ڋ׭‬ਏ݀ၨൂᐱ٬Ҕ CPT-11 ‫ܴ׳‬ᡉ 76Ƕ໳ೱનૈᡉ๱‫ڋ׭‬ϯᏢठ ᕎᏊ NDEA ‫ޑ‬ठᕎբҔǶ‫ݙ‬৔Α NDEA λႵ๏ϒ໳ೱનೀ౛ࡕǴဍ ዦวғ౗ᡉ๱फ़եǴ୏‫ނ‬ѳ֡ғӸਔ໔‫ ߏۯ‬77Ƕ. 5. ಒझຼය‫ޑ‬Ў᝘ӣ៝. 25.

(36) ! ! ಒझຼයǴࢂࡰૈ࡭ុϩ຋‫੿ޑ‬ਡಒझவ΋ԛԖํϩ຋่‫ࡕ״‬ғ ߏǴӆ‫ډ‬Π΋ԛϩ຋่‫ޑ״‬ൻᕉၸำǶಒझຼය‫ߏޑ‬อϸࢀΑಒझ‫܌‬ ೀ‫ރ‬ᄊǴ೭ࢂ΋ঁಒझ‫ނ‬፦ᑈಕᆶಒझϩ຋‫ޑ‬ൻᕉၸำǶᕎᡂ‫ޑ‬ಒझ аϷ੝‫ۓ‬໘ࢤ‫ޑ‬खजಒझததԖ౦த‫ޑ‬ϩ຋ຼයǶӚԛಒझϩ຋ϐ໔ ‫ޑ‬໔ሜǴхࡴ G1ǵSǵG2ǵM ѤයǶಒझຼයፓ௓ࢂӧӚය‫ޑ‬௓‫ڋ‬ᗺ (check points)΢຾Չ‫ޑ‬ǶಒझຼයύӸӧ‫ঁٿ‬ख़ा‫ޑ‬௓‫ڋ‬ᗺǺ΋ঁՏ ‫ ܭ‬G1/S ҬࣚೀǴӧথ٢ᜪ୏‫ނ‬ಒझύᆀࣁज़‫ڋ‬ᗺ(restriction point)Ǵ ྽ G1 යϣ‫҅ޑ‬ፓ࿯Ӣનಕᑈၲ‫ډ‬፾྽‫ޑ‬ำࡋԶёаຫၸ R ᗺǴаࡕ ൩ό໪٩ᒘಒझѦߦғߏӢનԶ໩‫ׇֹ‬ԋ᏾ঁಒझຼයǹќ΋ঁՏ‫ܭ‬ G2/M ҬࣚೀǴѬӧࢉՅᡏϩ໒‫ک‬ಒझϩ຋߻ߥ᛾ࢉՅᡏ֡ϩ‫ޑ‬ᆒዴ ‫ ܄‬78Ƕ. ! ! Ҟ߻Γॺว౜‫ޑ‬ಒझຼයፓ௓ӢኧࡐӭǴёᘜયΟεᜪǺ Ȑ1ȑಒ झຼයન(cyclins)ǵ Ȑ2ȑಒझຼයન٩ᒘ‫܄‬ᐟ䁙(cyclin dependent kinases, CDKs)ǵȐ3ȑCDK ‫ڋ׭‬ೈқ(cyclin dependent kinase inhibitor proteins, CKIs)ǶCDKs ࢂಒझຼයፓ௓‫ޑ‬ख़ाᕉ࿯Ǵ‫ځ‬೯ၸ٬Πෞೈ қ (pRb Ϸ‫࣬ځ‬ᜢೈқ p107ǵp130) ‫ޑ‬ᕗለϯֹٰԋፓ࿯բҔ‫ޑ‬ǹ Cyclins ࢂ CDKs ‫҅ޑ‬ፓ࿯ӢનǴૈ௴୏ CDK ࢲ‫܄‬ǹCKIs ࢂॄፓ࿯ ӢનǴё‫ ڋ׭‬CDK ࢲ‫܄‬Ƕ౜வ CDKs ϷΠෞፓ࿯ೈқǵCyclins ‫ک‬ CKIs ΟБय़ឍॊಒझຼයፓ௓Ӣનӧಒझຼයፓ࿯ύ‫ޑ‬բҔ 79Ƕ! 26.

(37) (1).CDKs Ϸ‫ځ‬Πෞೈқ. ! ! Ҟ߻ςว౜‫ ޑ‬CDK ԿϿԖΐᅿǴϩձ‫ڮ‬Ӝࣁ CDC2 ( CDK1 )ǵ CDK2ɴCDK9ǴѬॺӧ୷Ӣ‫ׇ‬ӈ΢‫ޑ‬ӕྍ‫܄‬ଯǴຬၸ 40ʘǶࢲϯ‫ޑ‬ CDK х֖໽ϯ٥ൂՏ(CDK)‫ک‬΋ঁ҅ፓ٥ൂՏ(cyclin)Ǵ߻‫ޣ‬ᗋሡ΋ ঁߥӺ‫ ޑ‬Thr ූ୷΢‫ޑ‬ᕗለϯ‫ ک‬Tyr ූ୷΢‫ޑ‬ѐᕗለϯǶ. ! ! CDK ೏௴୏ࡕǴё٬‫ځ‬ΠෞೈқวғᕗለϯǶ҂ᕗለϯ‫ ޑ‬pRb ૈ୼่ӝᙯᒵӢન E2FǴ٠‫ ڋ׭‬E2F ௴୏ S ය࣬ᜢ୷Ӣ‫ޑ‬ᙯᒵǶᕗለ ϯ‫ ޑ‬pRb ‫ ܈‬pRb ૈᏤठ E2F ‫ޑ‬ෞᚆǶෞᚆ‫ ޑ‬E2F ᆶ DP-1 ‫׎‬ԋ౦ྍΒ ᆫᡏǴૈ୼่ӝ‫ ډ‬DNA ‫ޑ‬੝‫ۓ‬ՏᗺǴ௴୏ S ය࣬ᜢ୷ӢᙯᒵǴ٬ຼ යຫၸ R ᗺǴಒझջவ G1 ය຾Ε S ය 80Ƕ (2).Cyclins ! ! Ҟ߻ςว౜‫ ޑ‬Cyclins Ԗ 12 ᅿǴϩձ‫ڮ‬Ӝࣁ Cyclin AɴHǶਥᏵ ಒझຼය‫ޑ‬ਔ࣬Ǵϩࣁ G1ǵS ‫ ک‬M යаϷբҔۘ҂ܴዴ‫ ޑ‬CyclinsǶ Cyclins ‫ޑ‬ၸࡋ߄ၲёа௴୏ CDK ࢲ‫܄‬Ǵ೷ԋಒझຼයፓ࿯Ѩ௓ǴᏤ ठဍዦ‫׎ޑ‬ԋǴӢԜ Cyclins Ψ೏ᆀࣁᕎೈқǶ ! ! G1 ය‫ ޑ‬Cyclins хࡴ Cyclin CǵDǵE ฻ΟࠠǴ‫ځ‬ύࡕ‫ࢂࠠٿ‬Ь ा‫ ޑ‬CyclinsǶCyclin D ԿϿԖ D1ǵD2ǵD3 ฻Οঁ٥ࠠǴΟࠠ Cyclin D ‫ޑ‬਽୷ለ‫ׇ‬ӈӕྍ‫܄‬ၲ 53.1ʘɴ63.5ʘǴѬॺ‫ ޑ‬C ҃ᆄԖ΋ঁ൤֖থ. 27.

(38) ਽ለȐPȑǵ‫ك‬਽ለȐEȑǵϺо਽ለǵํ਽ለȐSȑ‫ک‬᝵਽ለȐTȑ‫ޑ‬ ූ୷‫ׇ‬ӈǴᆀϐࣁ PEST ‫ׇ‬ӈǴᆶೈқ፦‫ޑ‬फ़ှԖᜢǹѬॺ‫ ޑ‬N ҃ᆄ Ԗ΋ঁᆶࢌ٤ᙯϯೈқӵ SV40 ‫ ޑ‬T ‫ל‬চǵဏੰࢥ E1A ೈқǵ٢ᓐዦ ੰࢥ E7 ೈқӅӕ‫ׇ‬ӈ Leu-X-Cys-X-GluǴԜ‫ׇ‬ӈ΋ኬёᆶ pRb Ϸ pRb ࣬ᜢೈқ่ӝǴᒿࡕӧ CDK4/CDK6 ‫ޑ‬բҔΠǴ٬ pRb ᕗለϯǴவԶ ٬ಒझவ G1 ය຾Ε S යǶ΋٤ғߏӢન(growth factors, GFs)ёᇨᏤѬ ॺ‫߄ޑ‬ၲǴ΋ѿѐନ GFsǴCyclin D1 ‫ޑ‬Нྗ཮ِೲΠफ़ǴӢԜǴᆀѬ ࣁғߏӢનག‫ڙ‬Ꮤ(growth factor sensors)108Ƕӧ G1 යǴ๏҅த‫ޑ‬ԋᠼ ᆢಒझᡉ༾‫ݙ‬৔‫ ל‬Cyclin D1 ‫ל‬ᡏǴૈ୼ߔЗಒझ຾Ε S යǴՠӧௗ ߈ G1/S Ҭࣚೀਔ߾‫ݙ‬৔คਏǴ೭΋่݀߄ܴ Cyclin D1 ӧ G1 ‫ޑ‬ύఁ යวච๱ख़ाբҔǶCyclin E ӧ G1 ය‫߄ޑ‬ၲఁ‫ ܭ‬Cyclin DǴӧ G1/S Ҭࣚೀၲ‫ډ‬ଯঢ়Ǵ຾Ε S යࡕ೴ᅌΠफ़Ǵ๏ಒझᡉ༾‫ݙ‬৔‫ ל‬Cyclin E ‫ל‬ᡏǴёߔЗಒझ຾Ε S යǴӢԜ೏ᇡࣁࢂፓ࿯ G1/S යᙯඤ‫ޑ‬Ѹሡ ೈқǶಒझ຾Ε S යࡕǴCyclin E फ़ှǴ‫ک‬Ѭ่ӝ‫ ޑ‬CDK2 ೏ញ‫ܫ‬р ٰ‫ ک‬Cyclin A ่ӝ 81Ƕ ! ! S ය Cyclins хࡴ Cyclin AǶѬࢂ Cyclins ύനԐ೏ว౜‫ޑ‬Ǵӧ G1 ఁය‫ ܭ‬Cyclin E ϐࡕ߄ၲǴՠ‫ ډޔ‬S යᆶ CDK2 ่ӝࡕω೏௴୏Ƕ ๏ಒझᡉ༾‫ݙ‬৔‫ ל‬Cyclin A ‫ל‬ᡏ‫܈‬ϸက Cyclin A ૈ‫ڋ׭‬ಒझ DNA ‫ޑ‬ ӝԋǴ߄ܴ Cyclin A ӧ S යԖख़ाբҔǶќѦǴѬӧ G2 යᆶ M ය‫ک‬. 28.

(39) CDC2ȐCDK1ȑ่ӝǴёૈᆶԖํϩ຋ԖᜢǴӢԜǴCyclin A Ψ೏ᆀ ࣁ M ය Cyclins82Ƕ (3).CKIs ! ! 1992 ԃǴXiong ฻ӧගྡྷપ Cyclin-CDK ፄӝ‫ނ‬ਔǴள‫ډ‬ΑќѦ ΋٤ೈқ፦Ǵࡕٰ᛾ܴ‫ځ‬ύࢌ٤ೈқࢂ၀ፄӝ‫ڋ׭ޑނ‬Ӣηջ CKIsǶCKIs ӧಒझຼයύ‫ڀ‬Ԗॄ‫܄‬ፓ࿯բҔϷӭኧӧΓᜪဍዦύँ ᡂѨࢲ‫ޑ‬٣ჴǴගҢ‫ځ‬ёૈࢂ‫׭‬ᕎ୷ӢǶਥᏵѬॺ‫ׇ‬ӈӕྍ‫܄‬Ǵёϩ ࣁ‫ঁٿ‬௼Ǻಃ΋௼хࡴ p16ǵp15ǵp18 ‫ ک‬p19ǴѬॺ֡х֖Ѥԛᗕೈ қ(ankyrin)ख़ፄ่ᄬ‫ک‬੝౦‫ޑ‬ፓ௓ Cyclin D-CDK4 ‫ ܈‬Cyclin D-CDK6 ࢲ‫܄‬ǹಃΒ௼хࡴ p21Ǵp27 ‫ ک‬p57ǴѬॺ‫ڋ׭‬ӭᅿ Cyclin-CDK ࢲ‫܄‬Ƕ. ! ! p21 ୷Ӣ‫ۓ‬Տ‫ࢉܭ‬Յᡏ 6p21.2ǴΞᆀࣁ cip1 (CDK-interacting protein 1)Ǵwaf1 (wild-typep53-activated fragment 1)Ƕথ٢୏‫ނ‬ಒझ ύǴp21 ‫ޑ‬фૈԿϿԖ‫ٿ‬Бय़Ǻ΋ࢂ CDK ‫ڋ׭ޑ‬ӢኧǴќ΋ঁࢂૈ ่ӝ‫ڋ׭ک‬ቚ෗ಒझਡ‫ל‬চ(PCNA)ǴவԶ‫ ڋ׭‬DNA ‫ޑ‬ፄᇙǶp21 ‫ޑ‬ N ᆄ‫ ک‬C ᆄϩձᏼॄଆ೭‫ٿ‬Бय़‫ޑ‬բҔǶp21 ૈ‫ڋ׭‬ӭᅿ Cyclin-CDK ࢲ‫܄‬Ǵ௢ෳ‫ँځ‬ᡂёૈࢂဍዦ‫׎‬ԋ‫ޑ‬ЬाচӢϐ΋ǶฅԶҞ߻ନΑӧ ᆮાᕎύว౜΋ঁಖЗஏዸη‫ޑ‬ᗺँᡂѦǴۘ҂‫ډـ‬ᜢ‫ځܭ‬дဍዦಒ झύ p21 ୷Ӣँᡂ‫ޑ‬ൔᏤǶp21 ୷ӢӧᙯᒵНྗ΢Ьा‫ ڙ‬p53 ፓ࿯‫ک‬. 29.

(40) ௴୏ǴԶ p53 ୷ӢँᡂӧΓᜪဍዦύࠅࢂத‫ޑـ‬Ǵࡺ౜ᇡࣁ p21 ୷Ӣ ЬाѨࢲᐒ‫ڋ‬ᆶ p53 ѨࢲԖᜢ 83Ƕp53 ‫ڀ‬Ԗ‫ׇ‬ӈ஑΋‫܄‬ǴΨ൩ࢂѬ཮ ᆶ୷Ӣፓ௓Տ࿼‫ޑ‬੝ਸਡੳለ‫ׇ‬ӈᗖ่Ǵ೭٤୷ӢੋϷಒझຼය‫ޑ‬ፓ ࿯ 84 Ƕ. 6. ಒझ঒Ϋ‫ޑ‬Ў᝘ӣ៝ ! ! ಒझीჄ‫܄‬ԝΫ(programmed cell death)Ψᆀࣁಒझ঒Ϋ (Apoptosis)!Ǵ೭ঁӜຒྍԾ‫׆‬ᛪЎǴཀࡘࣁယТ‫޸܈‬ᛐவ޸ਲ਼௞ပǴ ೭ঁӷബ೷‫ ܭ‬2:83 ԃǴҔ‫ܭ‬ඔॊಒझԝΫ‫ޑ‬΋ᅿࠠԄǴӧፄᚇ‫ޑ‬ғ ‫سނ‬಍ύᆢ࡭ว‫ػ‬Ϸಒझ໔‫ڀۓࡡޑ‬Ԗ๊ჹ‫ޑ‬ख़ा‫܄‬ǶӢԜಒझ঒Ϋ ࢂ΋ᅿಒझ‫ڙ‬ᕉნ‫ڈ‬ᐟࡕǴӧ୷Ӣፓ௓ϐΠ‫܌‬ౢғ‫ޑ‬ԾฅԝΫ౜ຝǴ ࡺҭᆀϐࣁಒझำԄԝΫǶၸำύҗ੝‫ޑۓ‬ೈқ፦Ϸሇનፓ௓ൻ‫ׇ‬ᅌ ຾٬ಒझԝΫǶ҅தಒझ೿ᏱԖགᔈ‫س‬಍ٰགᔈಒझϣ‫܈‬Ѧٰ‫ૻޑ‬ ৲Ǵ྽ୀෳ‫ډ‬όճ‫҅ځ‬தғߏ‫ૻޑ‬৲ਔǴ൩ЇวำԄԝΫǶ࣬ჹ‫ܭ‬ಒ झᚯԝǴಒझ঒ΫࢂಒझЬ୏ჴࡼ‫ޑ‬Ƕঁᡏ‫ޑ‬วғȐdevelopmentȑǵ ғߏȐgrowthȑǵϩϯȐdirentationȑၸำύǴಒझ঒ΫԖշ‫ܭ‬ᆢ࡭ғ ‫ނ‬ᡏϣࡡ‫ޑۓ‬фҔ 85Ƕ྽ಒझᎁ‫ڙ‬໾্Ъค‫ݤ‬অൺਔ཮೏੃ྐаٛЗ ‫׳‬ε‫ޑ‬Ӓ্‫ޑ‬ౢғǴ‫ٯ‬ӵಒझӧ๋Ѧጕ(UV)ྣ৔‫܈‬ϯᏢᛰᏊբҔΠ཮ ౢғँᡂ຾ԶᏤठᕎੱ‫ޑ‬วғǴՠ࿶җಒझ঒Ϋ‫ޑ‬ၸำё෧ե‫ځ‬วғ 30.

(41) ᐒ౗ 86ǶӧΓᡏϣಒझ঒Ϋਔӧಒझ‫׎‬ᄊ΢‫ک‬ғϯ੝ቻวғ‫ׯ‬ᡂǴ೷ ԋಒझጢ‫ޑ‬ያᕭ(Zeiosisor rapid blebbing)‫ࢉک‬Յ፦ᐚᕭϷಒझਡ‫ޑ‬஝ ှ(nuclear collapse)Ǵಒझ፦ጢ਻‫ݰ‬ϯǵझϣ DNA ೏फ़ှǵಒझ཮೴ ᅌያᕭ(shrink)ᆶᆫ໣(condense)ǵᕗિㄽํữለ(PS)வझϣӛѦᙌ рǴՠझᏔᆶಒझ‫ޑ‬ጢϝᆢ࡭ֹ᏾Ǵӕਔಒझϩ຋ԋঁձ‫ޑ‬ಒझ঒Ϋ ᡏ(apoptotic body)Ǵ٠җᎃ߈ಒझ‫܈‬ѮᏘಒझ‫܌‬տᏘǴԖձ‫ܭ‬΋૓Ӣ Ѧ໾‫ֽ܈‬೽લՈ‫܌‬೷ԋ‫ޑ‬ಒझᚯԝ(necrosis)Ǵ٠ё෧Ͽᡏϣౢғቃਗ਼ ‫ޑ‬խࣝϸᔈǶӢค‫ੱݹ‬ϸᔈౢғǴࡺ΋૓ᇡࣁᇨวಒझ຾Չಒझ঒Ϋ ࢂ΋ᅿၨӳ‫לޑ‬ᕎᐒ‫ڋ‬ǴᙖԜ෧Ͽᛰ‫ނ‬ёૈЇଆ‫ޑ‬ୋբҔǶ‫ܭ‬ᡏѦ୻ ᎦӢคѮᏘಒझǴ঒Ϋಒझനಖϝ཮‫و‬ӛᚯԝǶӧಒझ঒ΫၸำύǴ ಒझᕭλǴDNA ೏ਡለϣϪ䁙फ़ှԋ 180bp-200bp ТࢤǴ(ёа೯ၸ Ꮙጤႝ‫ݚ‬᛾ܴ*ǴԶಒझᚯԝਔǴಒझဍ๞Ǵಒझጢ೏ઇᚯǴ೯೸‫܄‬ ‫ׯ‬ᡂǶಒझᏔණပ‫ډ‬ಒझ໔፦ǴሡाѮᏘಒझѐమନǴ่݀ࢂ၀ֽ೽ ಔᙃว‫ݹ‬Ƕ࣬Кଆಒझᚯԝ-ಒझ঒Ϋࢂ‫׳‬த‫ޑـ‬ಒझԝΫ‫׎‬Ԅ 87Ƕ!. 7. ಒझᙯ౽ᆶߟΕ‫ޑ‬Ў᝘ӣ៝ ! ! “ᙯ౽”΋ຒԾ1829ԃҗ‫ݤ‬୯Jo sesh Claude Recamcer ගрǴϐࡕ Γॺჹဍዦᙯ౽ᐒ౛‫زࣴޑ‬ᕇளࡐӭԖሽॶ‫ޑ‬ཥၗૻǶ೴ᅌᕕှᕎಒ झ‫ޑ‬ᙯ౽ቹៜ‫ݯ‬ᕍϷႣࡕǴ٠ჹ‫ܭ‬ᕎੱ‫ޑ‬ൺวԖख़ा‫ޑ‬ቹៜǶൾ‫܄‬ဍ. 31.

(42) ዦӧບᘐਔр౜ֽ೽రЃ่ᙯ౽‫ޑ‬К౗1/3 ~ 1/4ǴԶဍዦठԝΫ‫ޑ‬Ь ाচӢ൩ࢂဍዦಒझ‫ޑ‬ᙯ౽Ƕ ! ! ᕎಒझ੆ዎ‫ک‬ᙯ౽ၸำߚதፄᚇǴεठёϩࣁаΠ൳ঁၸำǺ (1)ဍዦՈᆅ‫׎ޑ‬ԋǶ (2)ᕎಒझவচว೽ՏಥပǶ (3)ᕎಒझᆶಒझѦ୷፦ǵ୷ۭጢวғᗹߕ٠फ़ှǶ (4)ಥပ‫ޑ‬ᕎಒझऀຫՈᆅᏛ຾Εൻᕉ٠‫ډ‬ၲᇻೀ೽ՏǶ (5)ӧ஌ว೽Տᝩុቚ෗‫׎‬ԋᙯ౽ੰ‫؞‬Ƕ ! ! ဍዦߟΕᆶᙯ౽‫؁‬ᡯനԐࢂҗ Liotta ‫܌‬ගр‫ޑ‬ℚߕǵफ़ှ‫ک‬౽୏ Ο‫؁‬ᡯଷᇥǴՠࢂᒿ๱߈ԃٰჹဍዦࣁՈᆅࣴ‫ޑز‬ుΕǴว౜ဍዦ‫ޑ‬ ᙯ౽ᆶ༾Ոᆅ‫׎ޑ‬ԋஏϪ࣬ᜢǶ΋٤Ў᝘ᡉҢǴဍዦோԖ‫ڀ‬ഢՈᆅ‫׎‬ ԋ߄౜ࠠϐࡕωૈᙯᡂԋൾ‫ڀک܄‬ഢᙯ౽‫ૈޑ‬ΚǴᇥܴ༾Ոᆅ‫׎ޑ‬ԋ ࢂဍዦߟΕ‫ک‬ᙯ౽‫ޑ‬ᜢᗖǶ ! ! ဍዦՈᆅ‫׎‬ԋ‫ޑ‬ၸำᜪ՟‫҅ܭ‬தՈᆅғԋ‫ޑ‬ғ౛ᐒᙯǴ྽ဍዦಒ झϩ‫ݜ‬ՈᆅғԋӢનϐࡕǴё‫ޔ‬ௗ‫ڈ‬ᐟߕ߈‫ޑ‬ϣҜಒझ‫ׯ‬ᡂ‫ރ׎‬Ǵ٠ ᎂ౽‫ډ‬ဍዦߕ߈Ǵ໒‫ۈ‬ϩϯ‫׎‬ԋՈᆅ٠Քᒿ๱୷፦फ़ှ䁙аઇᚯ୷ۭ ጢǴᎂ౽ࡕ‫ޑ‬ϣҜಒझόᘐ‫ޑ‬ቚ෗ϩϯǴӧ‫߻ځ‬ጔ‫ޑ‬೽ҽ࡭ុቚ෗ϩ ϯǴԶࡕጔ‫ޑ‬೽ҽ೴ᅌ‫׎‬ԋᆅ๚่ᄬǴനࡕ‫׎‬ԋཥғՈᆅ 88Ƕ ! ! ೚ӭࣴ‫ز‬ᡉҢǴߦ຾Ոᆅ‫׎‬ԋ‫ޑ‬ӢનԖࡐӭǴ‫ٯ‬ӵVEGFǵBFGFǵ. 32.

(43) TGFǵIL-8ǵMMPsϷՈλ݈ғԋ‫ڈ‬ᐟӢનǶ‫ځ‬ύǴՈᆅϣҜғߏӢ ηȐVascular endothelial growth factorǹVEGFȑࢂനԖදၹཀက‫ڈޑ‬ ᐟӢηǴVEGFࢂϣҜಒझ஑΋‫ߦޑ‬Ԗํϩ຋ӢનǴЬाբҔ‫ܭ‬ϣҜ ಒझ΢‫ޑ‬flkǵflt1ǵflt4฻3ঁ‫ڙ‬ᡏǴ΋Бय़ёߦ຾༾ՈᆅϣҜಒझቚ ෗ᎂ౽Ǵќ΋Бय़ёߦ຾༾Ոᆅ೯೸‫܄‬ቚуǶԖჴᡍᡉҢǴКၨൂՏ ᡏᑈဍዦՈᆅ‫߄ޑ‬य़ᑈǴVEGF߄౜‫ޑ‬ဍዦࢂό߄౜‫ޑޣ‬Ϥ७ǴԶ VEGF߄ၲ‫ޑ‬෧Ͽᡉ๱फ़եဍዦᡏᑈՈᆅஏࡋ‫ک‬೯೸‫܄‬຾Զ೷ԋᙯ౽ ૈΚफ़եǶ ! ! Ꮓᆅόӕဍዦ໔‫ޑ‬ଆӢǵ୷Ӣࠠǵ߄౜ࠠԖࡐεৡձǴ٠Ъࡐӭ ཥ‫زࣴޑ‬ᡉҢӧᙯ౽‫ޑ‬ၸำ౐ੋ‫ࡐډ‬ӭፄᚇ‫ޑ‬ϩηӢનǴฅԶဍዦว ғ੆ዎᙯ౽‫ޑ‬ၸำࠅԖӅӕ‫ޑ‬ϩηᐒ‫ڋ‬ǴѬॺ೏ߚத‫ڐ‬ፓ΋ठӦಔᙃ ଆٰаӅӕวචբҔǴဍዦวғ੆ዎᙯ౽‫ޑ‬ၸำЬाࢂࡰᕎಒझ‫ޑ‬ℚ ߕफ़ှϷಒझ౽Չ຾Ε༾Ոᆅύ‫ޑ‬ၸำǶ ! ! ಒझℚߕȐcell adhesionȑӧൾ‫܄‬ဍዦᙯ౽ၸำύ‫ڀ‬Ԗख़ाբҔǴ ಒझℚߕхࡴӕ፦‫܄‬ᗹߕᆶ౦፦‫܄‬ᗹߕ‫ٿ‬ᅿǴ߻‫ࣁޣ‬ᕎಒझᆶᕎಒझ ໔‫ޑ‬ℚߕǴࡕ‫ࣁޣ‬ᕎಒझᆶ୷፦ಒझ໔‫ޑ‬ℚߕǶဍዦಒझ໔‫ߕޑ‬๱෧ ১٬ဍዦಒझಥᚆᆶຼᎁಒझ‫ߕޑ‬๱ǴԜࣁဍዦ੆ዎջᙯ౽‫؁߃ޑ‬Ƕ ௗ๱Ǵဍዦಒझℚߕ‫୷ܭ‬፦‫ک‬ՈᆅϣҜಒझ٠຾Εൻᕉ‫س‬಍೷ԋᙯ ౽Ƕ. 33.

(44) ! ! ӧୖᆶઇᚯಒझѦ୷፦‫ޑ‬䁙ύǴ୷፦ߎឦೈқ䁙ࢂख़ा‫ޑ‬䁙ᜪǴ Ҟ߻ς‫ޕ‬Ԗ 20 ӭᅿ MMPsǴЬाϩᜪԖȐ1ȑ໔፦ጤচ䁙ᜪȐ2ȑܴ ጤ䁙ᜪȐ3ȑ୷፦ྋનᜪȐ4ȑጢᜪǶ‫ځ‬ύܴጤ䁙ᜪύ‫ ޑ‬MMP-2 ‫ک‬ MMP-9 ࢂࣴ‫ز‬ၨӭ‫ٿޑ‬ᅿǶҞ߻εໆࣴ‫ز‬᛾ჴಒझѦ୷፦ ECM ‫ޑ‬फ़ ှӧဍዦೱᙹ‫؁‬ᡯύࢂᜢᗖ‫؁‬ᡯǴԶ MMPs फ़ှ ECM ‫ޑ‬ᐒ‫ࢂڋ‬Ӣࣁ MMPs ৎ௼ࢂ΋ঁೈқНှ䁙ৎ௼Ǵૈफ़ှӚᜪ‫ޑ‬ጤচǵᠼᆢℚೱೈ қǵቫℚೱೈқǵቸ‫܄‬ೈқ฻೭٤୷ۭጢମࢎǶќѦǴMMPs Ψૈග ଯϣҜಒझࢲ୏‫܄‬Ϸፓ࿯ᗹߕբҔǶ ! ! ဍዦಒझ‫ޑ‬ᑈཱུࢲ୏ቹៜ๱ဍዦಒझ຾Ε‫܈‬౽рՈᆅ‫ޑ‬ᚈӛࢲ ୏Ƕᕎಒझவ୷ۭጢઇཞೀෞᚆр‫ޑ‬БԄࢂаߓԯЃၮ୏‫ޑ‬БԄǶന ߈ว౜ဍዦಒझϩ‫ޑݜ‬ಒझᐟ䁙ёቹៜಒझ‫ޑ‬౽୏Ǵ΋٤୷፦ԋҽℚ ೱೈқǵᠼೱೈқϷࢌ٤ғߏӢηǴӵી৞નኬғߏӢηჹᕎಒझ‫ޑ‬ ࢲ୏‫ڀ‬ԖᖿϯբҔǴᔅշᕎಒझऀၸ୷ۭጢࡕǴ຾΋‫؁‬ྋှ໔፦‫่ޑ‬ ጌಔᙃӧ໔፦ύ౽୏Ǵ٠Ъӧၲ‫ډ‬ՈᆅᏛࡕёаҔӕኬ‫ޑ‬БԄ຾ΕՈ ᆅύǶ ! ! ߟΕ(invasion)ࢂࡰൾ‫܄‬ဍዦೱុόᘐӦ੆ዎǵઇᚯ‫ڬ‬ൎಔᙃᏔ‫۔‬ ‫ޑ‬ғߏ‫ރ‬ᄊǴջ‫ޔ‬ௗጭ‫ۯ‬Ƕᙯ౽ (metastasis)ࢂࡰൾ‫܄‬ဍዦಒझவচ ว೽ՏߟΕరЃᆅǵՈᆅ‫܈‬ᡏ๚Ǵᎂழ‫ډ‬дೀԶᝩុғߏǴ‫׎‬ԋᆶচ วዦӕኬᜪࠠ‫ޑ‬ဍዦǴ೭ঁၸำᆀࣁᙯ౽ǶဍዦಒझߟΕᆶᙯ౽ࢂ΋. 34.

(45) ঁፄᚇ‫ޑ‬ၸำǴ౐ੋ‫ډ‬೚ӭ‫ޑ‬ғߏӢηǵಒझӢન䁙‫س‬಍ϐ໔࣬ϕբ ҔԶֹԋ‫ ޑ‬89Ƕ ! ! ᙯ౽౜ຝ೯தࢂဍዦว৖‫ࡕޑ‬ය߄౜ǴӧΓᜪᕎੱύǴੰΓԝΫ ‫ޑ‬চӢຬၸԭϩϐΐΜǴࢂҗ‫ܭ‬ဍዦಒझᙯ౽‫يډ‬ᡏ‫ځ‬дᇿᇻ‫ޑ‬೽ ՏǴচว‫ޑ‬ဍዦࡐϿ཮೷ԋԝΫǶӧ΢Ҝಒझ‫ޑ‬ဍዦύǴᕎಒझ΋‫ۓ‬ ाऀၸ҅த΢Ҝಔᙃۭቫ‫ۭ୷ޑ‬ጢȐbasement membraneȑ΋ѿဍዦ ӧ୷ۭጢΠय़‫่ޑ‬ጌಔᙃ՞ளਥᏵӦϐࡕǴߏрٰ‫ޑ‬ဍዦಒझი཮ϩ ᚆрٰǴᅖΕߕ߈‫ޑ‬Ոᆅ‫ޣ܈‬రЃᆅ္Ǵ೭ᅿவ‫ڬ‬ൎಔᙃ຾ΕՈᆅ‫܈‬ ‫ޣ‬రЃᆅ‫ޑ‬ၸำǴᆀࣁϣᅖȐintravasationȑ Ƕ࿶ၸൻᕉ‫س‬಍‫܈‬రЃ‫س‬ ಍ǴՈᆅ‫܈‬రЃᆅஒဍዦಒझ஥‫ډ‬ᇻБ೽ՏǴ‫܈‬೚ဍዦಒझ൩ьӧࢌ ೀǴဍዦಒझаѦᅖȐextravasationȑ‫ޑ‬БԄᚆ໒Ոᆅ‫܈‬రЃᆅࡕǴ ൩ӧόӕ‫ޑ‬ಔᙃᏔ‫۔‬ԋߏǴߡ‫׎‬ԋᙯ౽ 90Ƕ. 35.

(46) ಃΟക 1.. ‫׷‬਑ᆶБ‫ݤ‬. ࣴ‫ز‬೛ी. ! ! ҁࣴ‫ز‬Ьाࢂ௖૸໳ೱનࢂցૈ୼‫ڋ׭‬ΓᜪՁᕎ SCC-4 ಒझਲ਼ ‫ޑ‬ғߏǴϷಒझ঒Ϋၡ৩‫ޑ‬ϩηբҔᐒᙯᆶ‫ڋ׭‬ΓᜪՁᕎ SCC-4 ಒ झਲ਼‫ޑ‬ᙯ౽ᆶߟΕբҔᐒᙯϐࣴ‫ز‬Ǵ٠຾΋‫زࣴ؁‬໳ೱનჹΓᜪՁᕎ SCC-4 ဍዦӧᇘႵࢲᡏϣ‫ޑ‬ϩηբҔᐒᙯϐࣴ‫ز‬ǴаΠࣁҁჴᡍ‫ࣴޑ‬ ‫ز‬೛ीࢎᄬȐӵკ 3-1ȑǶ. ΓᜪՁᕎTDD.5ಒझਲ਼ аόӕᐚࡋϐ໳ೱન)cfscfsjof*ೀ౛όӕ‫ޑ‬ਔ໔. ࢬԄಒझሺϩ‫݋‬. ችࢃ၂ᡍ. EBQJࢉՅ. ಒझᙯ౽ჴᡍ. ಒझӸ౗ࢲϩ‫݋‬ ROSΕMMPΕCa2+. ౦ᅿҜ౽෗୏‫ނ‬ኳࠠ. ಒझຼයϩ‫݋‬. ౽୏၂ᡍ. ՋБᏀᗺ‫ݤ‬ϩ‫݋‬. კ3-1 ҁჴᡍϐࣴ‫ز‬೛ीࢎᄬ. 36. ऀ೸၂ᡍ.

(47) 2.. ಒझਲ਼ٰྍ. ! ! ҁჴᡍ‫܌‬٬Ҕϐ SCC-4 ಒझਲ਼ࣁΓᜪՁᕎಒझਲ਼Ȑ(human squamous cell carcinoma of tongue ; 55 ྃ,‫ )܄ت‬ǴҗཥԮ१ࠔπ཰ࣴ ‫܌ز‬ȐFood Industry Research and Development InstituteȑᖼளǶ. 3. ᛰࠔ၂Ꮚ (1).5-BIS(4-HYDROXY-3METHOXYPHENYL)-1,4-HEPTADIENE-3,5 -DIONE (2).BerberineǺᖼԾ Sigma Chemical Co. (3).Dimethyl sulfoxideȐDMSOȑǺᖼԾ Sigma Chemical Co. (4).DMEM mediumǺᖼԾ Gibco (5).Fetal bovine serumȐजФՈమ, FBSȑǺᖼԾ Gibco (6).L-GlutaminȐᎽữለ , LGȑǺᖼԾ Gibco (7).Penicilllum StreptomycinȐPSȑǺᖼԾ Gibco (8).3,3’-Dihexyloxacarbocyanine iodide Ȑ DioC6 ȑǺ ᖼ Ծ Molecular Probes (9).propidium iodideȐPIȑǺᖼԾ Sigma Chemical Co. (10).Trypsin-EDTAǺᖼԾ Amersco (11).Trypan blueǺᖼԾ Sigma Chemical Co. (12).Disodium hydrogen phosphateȐNa2HPO4ȑǺᖼԾ Merck. 37.

(48) (13).Sodium chlorideȐNaClȑǺᖼԾ Merck (14).Potassium dihydrogen phosphateȐKH2PO4ȑǺᖼԾ Merck (15).Potassium chlorideȐKClȑǺᖼԾ Merck (16).Propidium iodideȐPIȑ ǺᖼԾ Sigma Chemical Co. (17).PhiPhiLux®-G1D1 kitǺᖼԾ OncoImmunin ȐGaithersburg, MD, USAȑ (18).RNase AȐRibonuclease AȑǺᖼԾ Ameresco (19).Triton X-100Ǻpurchase from Sigma chemical Co. (20).EthanolǺᖼԾ TEDIA (21).Ammonium persulfateȐAPSȑǺᖼԾ Amersco (22).Acrylamide/Bis 40 ʘ. solution Ȑ ACRYL/BISTM29:1 ȑǺ ᖼ Ծ. Amresco (23).Bovine serum albuminȐBSAȑǺᖼԾ Merck (24).GlycineǺᖼԾ Amresco (25).MethanolǺᖼԾ TEDIA (26).formaldehydeǺᖼԾ Merck (27).ECL kitȐEnhanced chemiluminescent kitȑ ǺᖼԾ Amersham (28).Protein assay-Dye reagent concentrateǺᖼԾ Bio-Rad (29).Protein markerǺᖼԾ Femantas (30).10X SDS bufferȐSodium dodecyl sulfateȑǺᖼԾ Amresco. 38.

(49) (31).TEMEDȐN,N,N’,N’-Tetramethyl-ethylenediamineȑ ǺᖼԾ Amresco (32).TrisȐTrisȐhydroxymethylȑ-aminomethaneȑǺᖼԾ Amresco (33).Tween 20ǺᖼԾ Amresco (34).ᡉቹᏊǺᖼԾ Kodak (35).‫ۓ‬ቹᏊǺᖼԾ Kodak (36).BioMax FlimǺᖼԾ Kodak (37).Agaros˧ǺᖼԾ Amresco (38).ਡለપϯ၂ᏊಔȐDNA purification kitȑ ǺᖼԾ Gene Mark (39).ೈқ፦๧‫ڗ‬၂ᏊȐprotein extraction solutionȑ ȐPRO-PREPȑ ǺᖼԾ iNtRON Biotechnology, INC. (40).5× TBE bufferǺᖼԾ Amresco (41).Primary antibody Ȑ1°‫ל‬ᡏȑ Ǻ (a). anti-actinǺᖼԾ Oncogen (b). anti-Bcl-2ǺᖼԾ upstate (c). anti-Bcl-Xl/XsǺᖼԾ upstate (d). anti-BaxǺᖼԾ upstate (e). anti-casoase-3ǺᖼԾ upstate (f). anti-caspase-7ǺᖼԾ Calbiochem (g). anti-caspase-8ǺᖼԾ upstate. 39.

(50) (h). anti-caspase-9ǺᖼԾ upstate (i). anti-Cytochrome CǺᖼԾ Oncogen (j). anti-cyclin EǺᖼԾ upstate (k). anti-cdk2ǺᖼԾ upstate (l). anti-cdk4ǺᖼԾ upstate (m).anti-cdk6ǺᖼԾ upstate (n). anti-Phosporylation ERKǺᖼԾ upstate (o). anti-JNKǺᖼԾ upstate (p). anti-Phosporylation JNKǺᖼԾ upstate (q). anti- NF kappa B Rel—a(p65)ǺᖼԾ upstate (r). anti-p38ǺᖼԾ upstate (s). anti-phosporylation p38 ᖼԾ upstate (t). anti-p53ǺᖼԾ Oncongen (u). anti-PARPǺᖼԾ upstate (42).Secondary antibodyȐ2°‫ל‬ᡏȑǺ (a). goat anti-mouse IgGȐHRPȑhorseradish peroxidase conjugated antibodyǺᖼԾ ChemiconǹAP124P (b).gout anti-rabbit IgGȐHRPȑhorseradish peroxidase conjugated antibodyǺᖼԾ Chemicon. 40.

(51) 4. ሺᏔ೛ഢǵᏔ‫׷‬ (1). Spectrolinker(UV box)ǺᖼԾդܳᑫ཰Ԗज़Ϧљ (2).ಒझ୻ᎦҝǺᖼԾ FALCON (3).ಒझ୻ᎦዬǺᖼԾ FALCON (4).ಒझ୻ᎦጃǺᖼԾ Nuaire (5).ಒझीኧᏔȐHaemocytometerȑǺᖼԾ Boeco (6).ॹҥԄՏႽৡᡉ༾᜔Ȑphase-contrast microscopeȑǺᖼԾ Olympus (7).༾ໆϺѳ (TE-200; MILLTER) (8).ѐᚆηНᇙ೷ᐒǺᖼԾ Minipore (9)ႝྍ‫ٮ‬ᔈᏔǺᖼԾ Amersham (10).ለᡵॶෳ‫ۓ‬ीȐC831ȑǺᖼԾ Consort (11).PVDF membraneǺᖼԾ Minipore (12).Mini-3D ShakerǺᖼԾ Boeco (13).SDS-PAGE ႝ‫ݚ‬ኲ঺ಔǺᖼԾ Bio-Rad (14).Transfer Cell Blot ঺ಔǺᖼԾ Bio-Rad (15).у዗݈ǺᖼԾ Lab-Line (16).ࢬԄಒझीኧሺȐFlow cytometryȑǺᖼԾ Becton Dickinson (17).ଯೲᚆЈᐒǺᖼԾ HERMLE (18).ϩӀӀࡋीǺfrom Beckman 41.

(52) (19).ӀᏢᡉ༾᜔ȐOlympus CH2ȑ (20).ӀᏢᡉ༾᜔ȐNikon LABOPHOT-2ȑ (21).ሇનխࣝϩ‫݋‬ሺȐanthos 2020ȑǺᖼԾ Anthos Labtec, Australia (22).DNA ႝ‫ݚ‬ኲǺᖼԾ Mupid-2. 5. ᛰࠔଛᇙ ໳ೱનȐBerberineȑપϯ‫ނ‬Ǵગ‫ ڗ‬37.18 mgǴྋ‫ ܭ‬10ml DMSOǴ ଛᇙԋ 10 mM stock solutionǴவ stock solution ϩձ‫ڗ‬όӕᐚࡋଛᇙ‫܌‬ ሡᐚࡋǴଛᇙᐚࡋӵΠǺ ϩηԄǺ[C20H18NO4]+ Molecular Weight = 371.8 ವԸᐚࡋȐMȑɨϩηໆ/ᡏᑈȐLȑ 371.8×103 mg / 1000 mlɨ1000 mM 37.18 mg / 10 mlɨ10 mM ! ! ಃ΋೽ҽӧ໳ೱનё೸ၸcaspase-8, -9‫ک‬-3 ࿶җAIFᆶEndoGၡ ৩ᇨวΓᜪՁᕎSCC-4ಒझਲ਼ಒझ঒Ϋ‫ޑ‬ϩηբҔᐒᙯϐࣴ‫ز‬ύǴஒ ଛᇙӳόӕᐚࡋ‫ޑ‬໳ೱનҔٰ຾ՉಒझӸࢲ౗၂ᡍȐViability assayȑ ٠ வ Ӹ ࢲ ౗ ၂ ᡍ ่ ݀ ύ ᒧ ‫ ڗ‬ъ ठ ԝ ᐚ ࡋ (IC50) Ǵ Զ IC50 ‫ ޑ‬໳ ጪ ન ȐBerberineȑᐚࡋ߾ࣁҁፕЎֹԋ࣬ᜢჴᡍ‫܌‬ҔϐᐚࡋǶҁჴᡍനಖ ᐚࡋϩձࣁ15 μMǵ25 μMǵ50 μMǵ75μMǵ100 μM(ӵ߄3-1)Ƕ 42.

(53) ߄ 3-1 ໳ೱનȐBerberineȑϐଛᇙ Concentration of Berberine. 10 mM of Berberine. DMSO. 15ȝ M. 150 ȝl. 850 ȝl. 25ȝM. 250 ȝl. 750 ȝl. 50 ȝM. 500 ȝl. 500 ȝl. 75 ȝM. 750 ȝl. 250 ȝl. 100 ȝM. 1000 ȝl. 0 ȝl. ! ! ಃΒ೽ҽӧ໳ೱન‫ڋ׭‬ΓᜪՁᕎSCC-4ಒझਲ਼೸ၸ FAK, IKK, NF-țB, u-PA ‫ ک‬MMP-2 ‫ ک‬-9 ၡ৩‫ޑ‬ಒझᙯ౽‫ߟک‬Ε‫ޑ‬ϩηբҔ ᐒᙯϐࣴ‫ز‬ύǴஒଛᇙӳόӕᐚࡋ‫ޑ‬໳ೱનȐBerberineȑҔٰ຾Չಒ झᙯ౽၂ᡍȐMigration assayȑಒझߟΕ၂ᡍȐInvasion assayȑ٠வ Ӹࢲ౗၂ᡍ่݀ύᒧ‫ڗ‬ъठԝᐚࡋ(IC50)ǴԶIC50‫ޑ‬໳ጪનȐBerberineȑ ᐚࡋ߾ࣁҁፕЎֹԋ࣬ᜢჴᡍ‫܌‬ҔϐᐚࡋǶҁჴᡍനಖᐚࡋϩձࣁ 62.5μMǵ125 μMǴଛᇙБ‫ݤ‬ӕ߄3-1Ƕ ! ! ಃΟ೽ҽӧ໳ೱનჹΓᜪՁᕎ SCC-4 ಒझਲ਼ӧᇘႵࢲᡏϣ‫ޑ‬ϩ ηբҔᐒᙯϐࣴ‫ز‬ύǴஒଛᇙӳόӕᐚࡋ‫ ޑ‬doxorubicin ᆶ Berberine Ҕٰ຾Չಒझ xenograft Model ၂ᡍǶҁჴᡍനಖᐚࡋϩձࣁ doxorubicin ࣁ 4 mg/kg ǵBerberine ࣁ 10 mg/kgǶ. 43.

(54) 6. ಒझ୻Ꭶ (1).୻Ꭶచҹ ! ! ΓᜪՁᕎ SCC-4 ಒझਲ਼а DMEN ୻Ꭶ୷బу 10 %जФՈమǵ5 % L-glutamine Ϸ 5 % Penicillin Streptomycin ϐ୻ᎦనǴ࿼‫ ܭ‬5 % CO2 Ϸ 95 % air Ϸྕࡋ 37ʚ‫ޑ‬ಒझ୻ᎦጃȐincubatorȑύ୻ᎦǴऊΒϺ‫׳‬ ඤ΋ԛ୻ᎦనԿಒझߏԿΖϩᅈࡕ߾ஒಒझ଺ᝩж୻Ꭶ‫܈‬ϩዬа‫ٮ‬ ჴᡍϐҔǶ (2).ᝩж୻Ꭶ ! ! ࡑಒझߏԿΖϩᅈਔǴӧค๵ᏹբѠϣஒ flansk ύ୻Ꭶన֎рҧ కǴӃа PBSȐphosphate buffer salineȑమࢱಒझኧԛǴࢱృࡕӆу Ε 3 ml trypsin ຾Ε‫؂‬౟ flansk ύǴ‫ܫ‬࿼‫ ܭ‬37ʚ୻Ꭶጃύೀ౛ 5 ϩដ ࡕ‫ڗ‬р٠ዴᇡಒझᅆੌ௃‫ݩ‬Ǵऩಒझε೽ϩςԋᅆੌ‫ރ‬ᄊӆуΕଳృ ୻Ꭶన 10 ml ຾Ε‫؂‬౟ flansk аύ‫ ک‬trypsin ϐբҔǴஒ౟ύ‫܌‬Ԗనᡏ ֎ԿᚆЈᆅύа 1500 rpm ᚆЈ 5 ϩដǴᚆЈࡕஒᚆЈᆅ‫ڗ‬ӣค๵ᏹ բѠϣ٠ॹ௞΢మనǴᇸ‫ܡ‬ᚆЈᆅஒಒझѺණӆуΕ 10 ml ཥᗲ୻Ꭶ నаႝ୏֎ໆᆅϸᙟ‫֎ܜ‬٬֡Ϭష‫ک‬Ǵ‫ ڗ‬20 Pl ϐಒझᝌੌనуΕ 80 Pl trypan blue ྋనǴаՈౚीኧዬȐCounting chamberȑीᆉಒझኧ ҞȐtrypan blue ཮຾Εಒझጢςઇཞ‫ޑ‬ಒझϣஒಒझࢉԋᙔՅǴԶֹ ᏾ಒझ߾ό೏ࢉՅȑǶ 44.

(55) ! ! ಒझኧीᆉǺF(ࣁ‫܌‬ीኧ‫ޑ‬ಒझኧ) u 10 (10 ml ୻Ꭶన) u 5 (trypan blue 5 ७ีញ)u 104 ٩ჴᡍҞ‫ޑ‬όӕǴीᆉ‫܌‬ሡಒझన࿼‫ܭ‬ό ӕ୻ᎦҝύǶ (3).ಒझհএߥӸ ! ! SCC-4 ಒझਲ਼࿶୻Ꭶ٠ዴᇡࠠᄊ҅தࡕǴёճҔనᄊේհএߥ ӸǴհএߥӸϐಒझᔈӧғߏ‫ܮ‬౰ЪӸࢲ౗ଯϐ‫ރ‬ᄊΠ຾ՉǶಒझհ এߥӸБ‫ࣁݤ‬Ǵհএ߻ᔈ‫ݙ‬ཀಒझғߏ௃‫׎‬Ǵ٠ӧ߻΋В‫׳‬ඤ୻Ꭶ ୷Ƕএಒझ߻ሡӃଛᇙհএߥӸྋనǺஒ DMSO Ȑdimethyl sulfoxideȑ уΕཥᗲ୻Ꭶ୷ύǴ٬‫ځ‬നಖᐚࡋࣁ 7ʘǴషӝ֡ϬǴ࿼‫ྕ࠻ܭ‬Πࡑ ҔǶஒाԏ໣೏հএϐಒझ٠ҔᚆЈᐒᚆЈಒझǴᚆЈࡕीኧಒझஏ ࡋϷಒझӸࢲ౗ǶᚆЈࡕॹ௞΢మనǴуΕ٣߻ଛᇙӳ‫ޑ‬հএߥӸ నǴ٬‫ځ‬ಒझኧӧ 2×106 ~5×106 cells/mlǴషӝ֡Ϭࡕ࿼‫ܭ‬а኱ҢӳӜ ᆀǵኧҞϷհএВයϐհএᆅύǴ1 ml / ᆅǶ ! ! հএߥӸБ‫ݤ‬Ǻհএᆅ࿼‫ ܭ‬4 °CǴ15 mino -20 °CǴ30 min o -80 °CǴ16~18 λਔoనᄊේኲߏයߥӸǶ (4).ᛰ‫ނ‬ೀ౛ ! ! ᝩжಒझֹӄຠᙟ౟ۭࡕǴ୻ᎦԿΖϩᅈਔߡёа೏ԏ໣Կ microplate ‫ ܈‬dish ϣ୻Ꭶа‫ٮ‬ჴᡍϐҔǴऩ‫ک‬ᛰ‫ނ‬Ӆӕ୻Ꭶόຬၸ 24 λਔ߾ёόሡ࿼ඤ୻ᎦనǴऩຬၸ 24 λਔ߾Ӄ࿼ඤԋཥᗲ୻Ꭶనӆ. 45.

(56) уΕᛰ‫୻ނ‬ᎦǶmicroplate ‫ ؂‬well уΕ୻Ꭶనᡏᑈ 2 mlǴ߾у 20 μl ᛰ‫ނ‬ǹ3 mlǴ߾у 30 μlǴ߾ᛰ‫ނ‬ᐚࡋ೏ีញԋচٰ‫ ޑ‬0.01 ७ࣁനಖ ᐚࡋǶ. 7. ࢬԄಒझሺӸࢲ౗ϩ‫݋‬ෳ‫ۓ‬ȐViability assayȑ ! ! ࢬԄಒझሺቶ‫ޑݱ‬ᔈҔ‫ܭ‬ᕎಒझᖏ‫׉‬ኬࠔᔠෳ྽ύǴࢂဍዦғ‫ނ‬ Ꮲࣴ‫ޑز‬ख़ाπ‫ڀ‬ϐ΋Ǵࣁᕎಒझ‫ޑ‬Ԑය‫ޑ‬ᛰਏຑ՗ග‫ٮ‬ख़ाၗૻǶ ࢬԄಒझϩ‫ޑೌמ݋‬ว৖ග‫ٮ‬΋‫ז‬ೲᔠෳЪё᎞‫ޑ‬Б‫ۓݤ‬ໆಒझᝌ ੌనǴऩଛӝ፾྽‫ࢉޑ‬ᏊǴ߾ёаჹᛰ‫ނ‬ೀ౛ಒझࡕ଺όӕ‫ޑ‬ਏ݀ຑ ՗Ǵ‫ڀ‬ԖྗዴϷ‫ז‬ೲ‫ޑ‬ᓬᗺǶ ! ! Propidium iodineȐPIȑࢂ΋ᅿਡለࢉᏊǴࢉᏊϐಔԋ(ӵ߄ 3-2)Ǵ ྽ಒझԝΫਔ཮຾Չᚯԝ‫܈‬ಒझ঒Ϋ‫ޑ‬ၡ৩ǴԝΫ‫ޑ‬ಒझጢ཮Ѩѐֹ ᏾‫܄‬Ǵ٬ள PI ё຾Εಒझϣ‫ک‬ਡለ่ӝǴPI ཮ᆶ DNA ᚈިᖥ௽ύ ϐ A=TǵCŁG ᗖ่‫ޑ‬ణᗖȐhydro bondȑௗӧ΋ଆǹӸࢲ‫ޑ‬ಒझӢ‫ځ‬ ಒझጢֹ᏾аठ PI ค‫کݤ‬ಒझϣ‫ޑ‬ਡለ่ӝǴ࿶ PI ࢉՅֹԋ‫ޑ‬ಒझ ёҗࢬԄಒझሺ‫ ܭ‬488 nm ‫ޑ‬ႜ৔ӀᐟวǴࢉ΢ PI ‫ޑ‬ԝΫಒझ࿶ႜ৔ Ӏᐟว཮և౜ၨமआՅᑻӀǴӸࢲಒझ߾คǴԝࢲಒझ٩ԜࣁϩᒣǴ ԶࢬԄಒझሺϣ೽ࢂа CellQuest ೬ᡏϩ‫݋‬ಒझӸࢲ౗Ƕ ! ! ჴᡍ‫؁‬ᡯࢂஒಒझ୻Ꭶ‫ ܭ‬6 well plate ύǴಒझ٩ྣ 2×105/well ᅿ෌‫୻ܭ‬ᎦҝύǴӆஒ୻Ꭶҝ‫ܫ‬࿼୻ᎦጃϣǴ‫ ؂‬well medium ᕴໆࣁ 46.